Leica Microsystems
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5th Oct, 2022

Considerations for multiplex live cell imaging

Multiplexing, or multicolor microscopy, in live cell imaging allows the observation of multiple cell structures and processes in synchrony, providing more physiologically relevant results.
Download this eBook to discover some key aspects of setting up a successful simultaneous multicolor live cell experiment as well as avoiding some of the major pitfalls. Plus, find out how the Mica imaging Microhub is highly adapted for live cell imaging experiments.
Learn more about:
Selecting the right fluorophore combinations
Avoiding the misleading effects of fluorescence crosstalk
Maintaining near physiological conditions
Considerations for image acquisition
12th Sep, 2022

Review Article – How to improve your confocal microscopy images

Learn about the common causes of autofluorescence and how to remove them from your confocal microscopy preparations. Depending on your application, there can be any number of sources of autofluorescence, but luckily there are just as many solutions—from changing your media to using lifetime imaging and far-red dyes.
Learn more about autofluorescence:
Possible causes and sources
Possibilities and advanced methods of elimination
Omission by experimental design
28th Jul, 2022

3D Tissue Imaging: From Fast Overview To High Resolution With One Click

3D Tissue imaging is a widespread discipline in the life sciences. Researchers use it to reveal detailed information of tissue composition and integrity, to make conclusions from experimental manipulations, or to compare healthy and non-healthy individuals.
In this article, you will learn how Mica helps researchers do 3D acquisition and AI-based quantification in one instrument with the world’s first Microhub.
Download this article and learn how!
18th Jul, 2022

eBook | Microscopy for Neuroscience Research

To investigate the structure and function of the nervous system at a cellular level, a microscope is often needed to analyse thick, complex specimens. This presents many challenges, including the problem of out-of-focus light. Download this comprehensive new eBook to see how you can overcome these challenges using THUNDER Imagers and AI-based analysis.
Learn more about:
Microscopy techniques and applications for neuroscience
Common imaging challenges and considerations
Real-life neuroscience case studies
THUNDER Imagers and Aivia AI analysis software
1st Jun, 2022

Fast, high aquity imaging and AI-assisted analysis of retinal cell damage

When imaging thick sections or whole-mounted samples using conventional widefield imaging, out-of-focus blur can limit the ability to quantify cellular and molecular features. Resulting from indiscriminate sample illumination, blur causes poor contrast of fluorophore channels, masks image information, and reduces the ability for clear and reproducible analysis.
Read this article to learn about:
Challenges of out-of-focus blur in widefield imaging
Creating clear, high contrast images of the cellular integrity of whole mounted thick samples
Generate 3D images on large samples in high speed with no post-processing
Automated AI-based object detection with high confidence
22nd Apr, 2022

Live Imaging Guide – Explore what is possible in confocal microscopy

Download this new comprehensive guide for live imaging applications for information on new labelling strategies, optimal imaging parameters, and the microscopy approach to take for your experiment. This article provides information on
New, less invasive, more physiological labelling strategies
Choosing the most appropriate imaging technology
Imaging parameters that keep specimens healthy throughout the experiment
Explore what is possible in live imaging with the STELLARIS platform.
9th Nov, 2021

Lithium-ion battery preparation for SEM analysis

Here we discuss details of lithium-ion battery preparation for SEM analysis and how to eliminate the artifacts from mechanical preparation by broad ion beam milling.
In this application note, you will learn:
Step by Step preparation of lithium batteries for electron microscopy analysis
A simple & fast approach for sample mounting
Efficient mechanical pre-treatment steps
The ion milling process and SEM analysis
Ready to discover how this works and what it could do for you?
28th Sep, 2021

App Note: Intravital imaging in a confetti breast cancer mouse model

Imaging possibilities for intravital breast cancer research
The ability to study cancer cell behavior in intact tissues within a live organism is crucial to gain biological insights into disease progression and to develop effective treatments for cancer.
Here we present to you an experimental approach that enables all fluorophores to be distinguished in such a complex sample, especially where they spectrally overlap.
In this application note, we discuss
The labeling method called lineage tracing
Why multiphoton microscopy is the most suitable imaging technique to image multicolor reporter lines
Why STELLARIS 8 DIVE with 4Tune is the ideal tool for deep in vivo multicolor imaging
Read our step-by-step guide to imaging confetti mice and the benefits of multiphoton confocal imaging in cancer research.
16th Sep, 2021

Download Free E-Guide: How to keep particulate contamination under control

Find out how to quickly identify particle contamination in medicine production
Identifying and eliminating particulate contamination in pharmaceutical products is often complex, time-consuming, and requires the particles to be relocated.
An innovative 2-in-1 method for materials analysis makes visual and chemical particle analysis faster, easier, and more cost-effective.
Find out about…
The risks and difficulties caused by particulate contamination of pharmaceutical products and how to keep them under control
How you can determine the shape, size, colour, and composition of particles more efficiently, reliably, and cost-effectively
How to image and analyze your samples independently of the composition without the need for time-consuming sample preparation or sample transfer between multiple instruments
The benefits of simultaneous visual and chemical particle analysis using the Leica DM6 M LIBS over conventional SEM/EDS root cause analysis workflows
Download this E-Guide and learn more about the Leica DM6 M LIBS and how your material analysis for contamination identification and root cause analysis can benefit.
26th Jul, 2021

Cryo-EM Sample Prep Masterclass - Curated tips and tutorials from the experts

Cryo-electron microscopy (cryo-EM) allows the visualization of biomolecules, from proteins to viruses, and allows snapshots of cellular processes and protein interactions to be seen at high resolution.
If you want to get the best results possible from cryo-EM you need to ensure you optimize the critical step of sample preparation.
To help you achieve reliable and robust results in your cryo-EM workflows, we have collected and collated a selection of tools (AppNotes, Webinars, Tutorials) featuring different techniques and aspects of cryo-EM sample preparation.
These include:
Key steps and instrumentation of cryo sample prep including vitrification, light microscopy screening, sectioning, planing, fracturing, milling, coating, and transfer.
How you can explore a particular time point of a cellular process at high resolution using optogenetics or electrical field stimulation in combination with HPF.
The importance of and strategies for keeping samples under cryo conditions throughout sample preparation workflows.
How high pressure freezing allows millisecond freezing of larger samples.
An understanding of how computational clearing improves the image quality of vitrified samples.
And more...
1st Jul, 2021

A New Method to Remove Out-of-Focus Blur

Fluorescence widefield microscopy can often result in image haze, obscuring features in the image that you are really interested in. We commonly refer to this as background noise or out-of-focus blur. You can remove this by background subtraction, but a better way to do this is with an innovative Leica opto-digital technique called Computational Clearing.
Computational Clearing uses an iterative algorithm to clear the out-of-focus haze while keeping the in-focus signals intact.
It doesn’t “generate” the desired image, but simply unmasks it by eliminating the unwanted background signals in the sample.
This method is a cost-effective and speedy alternative to methods such as structured illumination or spinning disc systems.
Discover how Computational Clearing can help you elucidate more details in your microscopy samples.
This concise report gives insights on the new technology.
17th May, 2021

Download eBook: Workflow Solutions for Industrial Research

EM Sample Preparation for Nano Technology
In this workflow booklet you will find a compilation of workflow solutions frequently used to conduct material sample preparation. Every workflow shows the relevant preparation step, the corresponding instrument that can be used and the resulting application image. You will also find links to useful further information regarding material sample preparation.
Solid state sample preparation
Polymer/rubbers sample preparation
Electronic materials and semiconductors
Metal, Ceramics Material, Hard Material Composites Processing
Vacuum transfer
If you have special requirements for your workflow or any questions regarding the topics shown here, our Leica experts will be happy to assist you at any time.
6th May, 2021

Download eBook: Guides to Confocal Imaging

Get closer to the truth – faster. Imaging beyond the diffraction limit
STED super-resolution allows you to study multiple dynamic events simultaneously, so you can investigate molecular relationships and mechanisms within the cellular context.
Here are 3 practical application notes giving both an overview of the various cutting-edge implementations of STED nanoscopy and clear guidelines on successful STED imaging.
1. The Guide to STED Sample Preparation
The secrets behind successful STED imaging
Which fluorescent labels can be used for single color STED imaging
The right protocol to obtain high quality images, with a high signal/noise (S/N) ratio, of interesting structures in a specimen.
2. Nanoscopy meets Lifetime: Introducing τ-STED
What’s behind the unique τ-STED functionality - new approach to STED based on fluorescence lifetime that delivers cutting-edge image quality and gentle live-cell conditions.
3. Extending Nanoscopy Possibilities with STED and exchangeable fluorophores
A new approach enables whole cell, 3D, multi-color STED imaging as well as live-cell STED microscopy
8th Oct, 2020

E-Book | The Guide to Live Cell Imaging

In life science research, live cell imaging is an indispensable tool to visualize cells in a state as in vivo as possible. This E-Book reviews a wide range of important considerations to take to ensure successful live cell imaging.
What’s inside:
What parameters need to be considered to maintain cell viability
A full review of the wide range of Live Cell Imaging Techniques and Applications
An insight into the problem of Out-of-Focus Light in Live Cell Microscopy and how to overcome it
An introduction to the THUNDER Imagers and the DMi8 S platform that are ideally suited to cover all your live cell imaging applications.
Download this booklet and learn how to make your live cell imaging more successful!
1st Oct, 2020

Application Booklet – Ion Beam Preparation of Samples for SEM

Today, ion beam milling is one of the most widely-used methods for preparing samples for electron microscopy. During this process, the sample material is bombarded with a high-energy argon-ion beam to achieve high quality cross-sections and polished surfaces whilst minimizing deformation or damage.
Did you know that it is possible to create high-quality cross-sections through a touchscreen to display all layers - polymer layer, layers of adhesive and glass substrate? Did you ever try to display the grain boundaries and substructures in the grains of synthetic rock salt?
Specialists from Leica Microsystems have collected over 20 different examples featuring different samples for SEM in one booklet:
Cross sectional sample preparation
Preparation of semiconductor material
Preparation of metals
Preparation of stones
Preparation of paper, wood, rubber
Preparation of thermally sensitive samples
Cleaning sample surfaces and contrast enhancement
Processing large sample surfaces with the rotary stage
Sample preparation for EBSD
9th Sep, 2020

Review Article - Uncover Hidden Dimensions in Research

Enhance your research with Lifetime Imaging.
The vast majority of imaging experiments measure fluorescence intensity but there is another key property of fluorescence—it’s lifetime, information that comes “for free” with every experiment.
In this review article you will find:
What lifetime imaging is.
How it can improve quality and add depth to your applications.
How FLIM reduces the chance of artifacts and improves contrast.
Why you will be able to distinguish more fluorophores.
Additional possibilities for functional imaging (Molecular interactions, Biosensing, Species Separation).
9th Sep, 2020

Get Closer to the Truth: STELLARIS - Confocal Re-Imagined

A new chapter in the history of Confocal
Watch the On-Demand Webinar: Stellaris‚ Confocal Re-Imagined to learn how:
To set up complex experiments in a few clicks
To obtain more data from your experiment
To improve image quality while preserving temporal resolution
To localize areas of importance and to identify relevant details through easy navigation
1st Sep, 2020

Application Note – Add an extra dimension to your confocal results TauSense: With integrated fluorescence lifetime information

Use lifetime-based information to explore the function of molecules within cellular context, Improve image quality or separate fluorescent species beyond spectral options.
Read the Application Note to find out how you can use TauSense technology to:
Explore a new dimension of information in every confocal experiment with instant access to functional information
Improve image quality by removing unwanted fluorescence signals that mask your data and prevent you from truly seeing your sample.
Separate fluorescent species based on lifetime information, allowing you to distinguish fluorescent signals with similar spectra.
1st Sep, 2020

Get Closer to the Truth: STELLARIS - Confocal Re-Imagined

A new chapter in the history of Confocal
Watch the On-Demand Webinar: Stellaris‚ Confocal Re-Imagined to learn how:
To access to the hidden fluorescence lifetime-based information of your fluorophores
To access functional information such as metabolic status, pH and ion concentration
To maximize detection efficiency by removing unwanted fluorescence while preserving the desired signal
To distinguish dyes with similar overlapping spectra
24th Aug, 2020

Get Closer to the Truth: STELLARIS - Confocal Re-Imagined

A new chapter in the history of Confocal
Watch the On-Demand Webinar: Stellaris‚ Confocal Re-Imagined to learn how:
To achieve higher photon detection efficiency and extremely low dark noise, to image with more brightness and more detail
To tune exicitation and detection to perfectly match the spectral profile of the dyes in your sample to obtain clear and sharp images
To acquire more accurate and reliable data to proof your hypotheisis
19th Aug, 2020

Article – Beyond the rainbow: How to multiplex beyond the spectral limitations in multicolour confocal microscopy

Good spectral separation of fluorophores in multicolor confocal microscopy is fundamental to achieving high quality results and being able to include more fluorescent probes in your experiment.
In this review article we explore:
Common challenges in spectral separation
Several strategies you can take to improve the separation of fluorophores
An alternative technology - Fluorescence Lifetime imaging to separate fluorophores with overlapping spectra
9th Jun, 2020

THUNDER Imagers: High Performance, Versatility and Ease-of-Use for your Everyday Imaging Workflows

Free On-Demand Webinar
Dr. Remco Megens, PI at the Institute for Cardiovascular Prevention (IPEK) of the Ludwig-Maximilians-University in Munich, Germany
Dr. Falco Krüger, Advanced Workflow Specialist Widefield, Life Science Research Division, Leica Microsystems
THUNDER Imagers are a brand-new class of widefield microscopes developed by Leica Microsystems. They can acquire imaging data, even of thick 3-dimensional specimens, with high quality and speed.
However, there is more to life science experiments than beautiful images. Robust quantification and repeatability of imaging experiments are crucial. THUNDER Imagers provide the full workflow, from image acquisition to data analysis, with an amazing ease of use.
This webinar will showcase the versatility and performance of THUNDER Imagers in many different life science applications: from counting nuclei in retina sections and RNA molecules in cancer tissue sections to monitoring calcium waves in Arabidopsis seedlings and much more.
26th May, 2020

Download Your Personal EM Sample Preparation Workflow Guide

Our Leica specialists know that you are very busy working in the laboratory. So to save you time, they compiled a booklet with the details of appropriate workflows for the most frequently used EM specimen preparation methods. In the booklet you will find information on workflows for:
Correlative Methodologies
Surface Analysis
3D & 2D Tissue & Cellular Morphology
Suspensions & Macromolecules
The workflows and solutions discussed in this 26 page booklet are proven to give rapid, reliable, and reproducible results. Download this booklet and make your life easier.
14th May, 2020

How to Sanitize a Microscope

Due to the current coronavirus pandemic, there are a lot of questions regarding decontamination methods of microscopes for safe usage. Microscopes are commonly shared by several users, so that is why they can have the risk of contamination with microorganisms. In addition, microorganisms themselves can serve as specimens which are observed with the microscope. Thus, a frequent sanitization of the microscope is highly recommended. To avoid infections, the use of disposable gloves for microscope operation, cleaning, and decontamination is recommended as well. Disposable gloves can be decontaminated with, e.g., alcohol, if necessary, or should be changed frequently to minimize the risk of contamination.
10th Mar, 2020

Key Factors for Efficient Cleanliness Analysis

This article gives an overview of factors which are essential for an efficient cleanliness analysis process concerning automotive and electronics manufacturing.
In the automotive and electronics industry, even the smallest particulate contamination on components or parts may influence their performance, causing them to malfunction or decrease their lifetime. For automobiles, the filter systems are particularly vulnerable. For electronics, contamination on printed circuit boards (PCBs) or connectors may cause short circuits. So, cleanliness plays a central role in quality control for modern manufacturing, especially when using components produced by various suppliers. Contamination on critical components for a vehicle or device can lead to a breakdown of the entire system. Therefore, an efficient cleanliness analysis process must begin at the supplier level.
The following points are addressed in this article:
Most important factors for efficient cleanliness analysis
Optimization of the overall analysis workflow
18th Feb, 2020

How to prepare samples for Stimulated Raman Scattering

Guidelines for sample preparation and workflows on SRS microscopy
Stimulated Raman Scattering (SRS) is an emerging microscopy technique that images the vibrations of chemical bonds within a specimen. It allows researchers to probe the endogenous chemistry or biochemistry of their specimen without the need for exogenous labels. Some common SRS applications can be found in the Coherent Raman Scattering Microscopy Publication List.
Here, we offer guidelines for sample preparation and suitable workflows with a focus on SRS spectroscopic imaging.
These guidelines are for users of the TCS SP8 CARS microscope with SRS Option. It is your versatile, hands-free turnkey solution for vibrational imaging from Leica Microsystems. It offers both SRS and CARS imaging modalities which are fully integrated into the SP8 visible confocal microscopy platform. In parallel to CARS and SRS, additional nonlinear optical signals, such as two-photon fluorescence (2PF) and second harmonic generation (SHG), can be detected.
8th Jan, 2020

Key Factors to Consider When Selecting a Stereo Microscope

Stereo microscopes are often nicknamed the “workhorse” of the lab or production site. Users spend many hours looking through the eyepieces inspecting, observing, documenting, or dissecting samples. Careful assessment of the relevant applications for which the stereo microscope is needed is key to long-lasting, satisfactory use. Decision makers need to be certain that they can tailor the instrument exactly to their requirements.
To help users with decision making when selecting a stereo microscope, the main factors to consider are highlighted here.
27th Nov, 2019

Multiphoton Microscopy

Chat with Dr. Thomas Mathivet about In Vivo Cardiovascular Imaging of the Brain
Dr. Thomas Mathivet, working at the Cardiovascular Institute in Paris, France, was interviewed by Dr. Luis Alvarez, Product Application Manager, during Leica Microsystems Meets Science 2019 in Mannheim, Germany. He discusses his research on the cardiovascular system in the brain. The goal is a better understanding of specific disease conditions, such as brain tumors. Dr. Mathivet’s team performs real-time imaging of the brain in live animals and the results play a significant role in the development of new intervention strategies for these pathologies. The SP8 DIVE spectrally tunable multiphoton microscope has changed the approach of his team to imaging. The need to visualize simultaneously with 5 to 6 different channels, not possible with traditional systems, is now met. Dr. Mathivet has found that the new imaging capabilities of the SP8 DIVE enable him to see more, including label-free visualization of collagen to study the brain’s structural aspects.
4th Nov, 2019

SP8 FALCON: a novel concept in fluorescence lifetime imaging enabling video-rate confocal FLIM

SP8 FALCON (FAst Lifetime CONtrast) is a fast and completely integrated fluorescence lifetime imaging microscopy (FLIM) confocal platform. SP8 FALCON delivers video-rate FLIM with pixel-by-pixel quantification, thanks to a novel concept for measuring fluorescence lifetimes built on fast electronics and sensitive spectral hybrid detectors. Photon arrival times are recorded at count rates typical for standard confocal imaging. The system has ultra-short dead time and powerful built-in algorithms for data acquisition and analysis. The deep integration of FLIM into the confocal platform provides easy access to complex FLIM experiments.
25th Sep, 2019

Introduction to Ultramicrotomy

Ultrathin sectioning of biological specimens and materials for microscopy and array tomography (AT)
When studying samples, to visualize their fine structure with nanometer scale resolution, most often electron microscopy is used. There are 2 types: scanning electron microscopy (SEM) which images the sample surface or transmission electron microscopy (TEM) which requires a very thin, electron-transparent sample. Thus, to image the fine structure inside a sample using electron microscopy, the solution is to make very thin sections of it. The sample preparation method known as ultramicrotomy can produce ultrathin sections (20 to 150 nm thick) with minimal artifacts. During sectioning, the block face of the sample, from which the sections are cut, always remains a flat and straight surface which can be investigated with SEM. When the sections are imaged in an array, then a 3D image can be reconstructed of the sample. This approach is called array tomography (AT). An overview of ultramicrotomy and its use for AT is given below.
12th Sep, 2019

The Guide to STED Sample Preparation

This guide is intended to help users optimize sample preparation for stimulated emission depletion (STED) nanoscopy, specifically when using the TCS SP8 STED 3X nanoscope from Leica Microsystems.
It gives an overview of fluorescent labels used for single color STED imaging and a ranking of their performance. Fluorescent label combinations for dual and triple color STED imaging that minimize cross-talk during detection are recommended. There is a discussion of considerations for immunofluorescence labeling and a detailed protocol to obtain high quality images, with a high signal/noise (S/N) ratio, of interesting structures in a specimen. Important details for sample mounting and substrates that enable optimal imaging, minimizing aberrations and autofluorescence due to the mounting medium, are reviewed. Finally, for STED imaging of live-cells, the most appropriate fluorescent labels are mentioned, both fluorescent proteins (FPs) and organic fluorophores which give the best performance.
27th Aug, 2019

How FRET/FLIM Biosensors and Lifetime Imaging Helped Identify the Importance of Zinc for Human B-Cell Development

Interview with Dr. Mukta Deobagkar
In her latest studies, Dr. Mukta Deobagkar-Lele investigated the role of ZIP7, a zinc transporter protein, in modulating B cell activity and its effects on human immunity. During an interview with Dr. Luis Alvarez from Leica Microsystems, she describes the key findings of these studies published in her recent article in Nature Immunology. She also discusses her introduction to lifetime imaging with the SP8 FALCON and how this modality helps her explore B-cell biology and related immunodeficiencies.
22nd Jul, 2019

Expert Knowledge on High Pressure Freezing and Freeze Fracturing in the Cryo SEM Workflow

An interview with Roland A. Fleck, Director of the Centre for Ultrastructural Imaging at King’s College, London.
The Centre for Ultrastructural Imaging at King’s College London, UK are focusing their research on life sciences, specifically on neurobiological topics i.e. the organization of the brain and how synaptic vesicles fuse at membranes. They also support related medical studies, like malarial parasite projects or chemotherapy and behaviour, in particular animal cell model systems and development. Get an insight in the working methods of the laboratory and learn about the advantages of Cryo SEM investigation in EM Sample Preparation. Find out how high pressure freezing, freeze fracturing and cryo transfer add to the Cryo SEM workflow and how the Leica portfolio ensures the compatibility between these different steps.
10th Jul, 2019

Alzheimer Plaques: fast Visualization in Thick Sections

The Shamloo lab used Leica Microsystems novel THUNDER Imager 3D Cell Culture to acquire large Z-stacks of uncleared, 40μm-thick mouse brain tissues.
More than 60% of all diagnosed cases of dementia are attributed to Alzheimer’s disease. Typical of this disease are histological alterations in the brain tissue. So far, there is no cure for this disease. A few treatments try to slow down the fatal path or try to relieve the patients from symptoms. The laboratory of Dr. Mehrdad Shamloo at Stanford University studies pathological brain function with the goal of contributing to the discovery of novel therapeutics for Alzheimer’s disease. They use a mouse model of this disease to study the role of inflammation in Alzheimer’s disease progression. This requires imaging of thick uncleared brain tissues.
18th Jun, 2019

Visualize mechanical interactions of Cancer Cells

Cells interact with their environment. Not only on the base of chemical signals, but also by sensing and modifying mechanical properties of the extracellular matrix. The research goal of Dr. Bo Sun’s group at Oregon State University is to understand such interactions. One of their tools is visualization of matrix properties with a TCS SPE confocal microscope.
23rd May, 2019

Bridging Structure and Dynamics at the Nanoscale through Optogenetics and Electrical Stimulation

Part 1: The overall workflow for unravelling processes at the nanoscale in a practical way
Nanoscale ultrastructural information is typically obtained by means of static imaging of a fixed and processed specimen. However, this is only a snapshot of one moment within a dynamic system in which structures are constantly changing. Exploring specific time points of a dynamic process is therefore a major challenge. Exploring a process at the nanoscale through optogenetics or electrical field stimulation in combination with timed millisecond precision vitrification is a promising technology to overcome this challenge.
In the first part of a series of application notes the practical considerations of stimulation-assisted vitrification are discussed.
17th May, 2019

Phasor Analysis for FLIM (Fluorescence Lifetime Imaging Microscopy)

The phasor analysis approach to analyze fluorescence lifetime does not require any fitting. Phasor FLIM (fluorescence lifetime imaging microscopy) provides a 2D graphical view of lifetime distributions. This graphical view enables any observer to distinguish and separate different lifetime populations within a FLIM image rapidly. The interpretation of phasor FLIM distributions is straightforward. Multiple molecular species are resolved within a single pixel, because every species has a specific phasor.
26th Apr, 2019

Zebrafish Brain - Whole Organ Imaging at High Resolution

Structural information is key when one seeks to understand complex biological systems, and one of the most complex biological structures is the vertebrate central nervous system. To image a complete brain dissected from a developing zebrafish, one would need to cover a field of some ten square millimeters at a depth in the millimeter range.
Usually, low magnification lenses do not provide sufficient resolution to reveal the intricate structural interactions in nervous tissue. Additionally, due to scattering processes, the depth at which one can image within dense biological tissue using a confocal microscope is generally restricted to approximately 10 microns.
1st Apr, 2019

Streamline the workflow for wound healing assay

How can PAULA help streamline the workflow for my wound healing assay?
Application for wound healing assays
The wound healing assay, often used as a synonym for scratch assay or migration assay, is an important tool for the evaluation and measurement of cell migration, whether single cells or cell populations. Cell migration is tightly controlled during ontogenetic processes and at times can get out of control, e.g., during cancer development and progression.
Important questions to address concerning cell migration are:
How does a particular drug alter the migration rate of my cells?
How fast do my cells migrate under certain conditions?
11th Mar, 2019

THUNDER Technology Note

THUNDER Imagers: How Do They Really Work?
THUNDER Imagers deliver in real time images of 3D specimens with sharp contrast, free of the haze or out-of-focus blur typical of widefield systems. They can even image clearly places deep inside a thick specimen. Exactly how are they able to do this? This tech note explains how the THUNDER technology works.
The THUNDER Technology: Computational Clearing
THUNDER Imagers use an integrative holistic approach to overcome the limitations of camera-based imaging systems. They expand camera-based imaging to a new application range for 3D samples. This approach largely comes down to a new, innovative Leica opto-digital technique called Computational Clearing (CC).
Computational Clearing detects and removes the unwanted signals from out-of-focus regions of the specimen in real time. It clearly reveals the desired signals from the in-focus region of interest within the specimen. CC distinguishes between the out-of-focus and in-focus signals via the difference in size of the features. Together with the feature size, CC automatically takes all relevant optical parameters into account. CC starts in real time right at the beginning of the image acquisition.
The CC method can successfully visualize specimen details with sharp focus and contrast for specimens that are usually not suitable for imaging with standard widefield systems. Computational Clearing can be combined with image restoration methods.
THUNDER Imagers come with a number of different options in combination with Computational Clearing that are optimized for the various types of samples.
Download the full technology note
Interested to know more about how THUNDER technology works?
Please fill in the form to receive the full Technical Note.
26th Feb, 2019

Confluency Check with PAULA Cell Imager

Monitor your cell culture around the clock with PAULA and know the perfect time-point for your experiment
Many cell-based experiments require cells in a certain state. This can include their morphology, fluorescent protein expression, and the confluency of the cell layer. Researchers check their cells regularly to determine these attributes. Confluency estimation can be very difficult! This guess work is biased by every individual researcher, and can result in lab error.
Confluency Workflow
Read further to learn how PAULA – the Personal AUtomated Lab Assistant from Leica Microsystems – helps you to automate confluency checks and find the perfect time-point for your experiment.
28th Jan, 2019

Decode 3D biology in real-time with the new Leica imagers

THUNDER imager family enables users to decode 3D biology in real-time*
Wetzlar. World-leading designer and manufacturer of innovative microscope solutions, Leica Microsystems, has announced the launch of a new class of instruments for high-speed, high-quality imaging of a large diversity of samples, including thick specimens. THUNDER Imagers allow to visualize clearly in real time details even deep inside thick samples of biologically relevant models like model organisms, tissue sections, and 3D cell cultures. This makes THUNDER Imager a cost-effective and speedy alternative to methods like structured illumination or spinning disc.
THUNDER Imager eliminates the out-of-focus blur that clouds the view of thick samples when using camera-based fluorescence microscopes. This performance advantage is achieved with a new opto-digital method created by Leica Microsystems called Computational Clearing. Currently unique to the market, the new technology deployed in THUNDER Imagers makes visualization and analysis of large volume, thick specimens ideal for many biomedical applications where they are required, including regenerative medicine, cancer, and stem cell research to decode 3D biology in real time.
Our product managers talk about key advantages that the THUNDER Imager 3D Live Cell & 3D Cell Culture offer. Check our Video Here
Image - HeLa cell spheroid stained with Alexa Fluor 568 Phalloidin (Actin) and YOYO 1 iodide (Nucleus). Acquired with a standard wide field fluorescence microscope (left half) and an exposure of the same sample taken with a THUNDER imager (right half).
9th Jan, 2019

A Molecular Link between Cell Migration and Vascular Disease

Understanding the Factors that Control Healthy and Unhealthy Blood Vessel Development
Blood vessels transport vital nutrients and oxygen to all the cells in the body. Guided by a complex signaling network, endothelial cells sprout, proliferate, and migrate to form those vessels. One of the processes of vascular formation is angiogenesis, the growth of new vessels from pre-existing ones. Our current knowledge of the mechanisms controlling angiogenesis and vascular stability is still limited. However, these mechanisms play important roles in the pathophysiology of cancer, diabetes and stroke. Understanding the factors that control healthy and unhealthy blood vessel development is fundamental to prevent and fight such diseases.
In a recent study, researchers from Uppsala University shed light onto one of the mechanisms controlling angiogenesis. They addressed the endothelial function of Cdc42, a protein known to regulate the actin cytoskeleton and cell polarity. Using conditional knockouts and multicolor fluorescence imaging in a mouse retina model, the authors found that Cdc42 ensures proper cell migration, whereas in its absence cells proliferate but fail to distribute properly, leading to severe vascular malformations. This work was published in the journal Development.
23rd Nov, 2018

The Live Cell Microscopy Masterclass Series

The Easy Way to Boost your Live Cell Microsopy Expertise
From Bitesize Bio and Leica Microsystems
This Masterclass delivers regular, bitesize training from world experts straight into your inbox.
No charge, no obligations. Just an easy way to build your knowledge over time.
What you can expect in the Masterclass:
- Video Tutorials
- How-to Articles
- eBooks
- Methods Papers
What others are saying...
Dr Martin Wilson, Microscopy Editor, Bitesize Bio
"If you are serious about improving your live cell imaging skills, this series is a must. It takes you into the labs of an array of experts and gives you the benefit of their hard-won experience. Just as importantly, because the materials are delivered to you in bitesize pieces, the learning is gradual and doesn't take up much of your precious bandwidth."
12th Nov, 2018

Image Gallery: Tunable Multicolor Deep In Vivo Imaging

With the SP8 DIVE Multiphoton Microscope
Today’s life science research focusses on complex biological processes, such as the causes of cancer and other human diseases. A deep look into tissues and living specimens is vital to understanding the conditions and mechanisms in cells and finding answers to crucial questions challenging the life sciences.
The SP8 DIVE Deep In Vivo Explorer is a multiphoton microscope with spectrally tunable non-descanned detection that provides unlimited flexibility, so that users can optimally distinguish components and structures deep in tissues and living organisms. The SP8 DIVE comes with the unique 4Tune detection that offers full spectral freedom and allows the detection of multiple transgenic markers with perfect color separation in a single experiment.
23rd Oct, 2018

On-Demand Webinar on Laser Microdissection

Live Cell Isolation by Laser Microdissection
Speaker: Dr. Oleg Podgorny, Koltzov Institute, Moscow, Russia
Laser microdissection is a tool for the isolation of homogenous cell populations from their native niches in tissues to downstream molecular assays. Beside its routine use for fixed tissue sections, laser microdissection may be applied for live cell isolation.
Unlike other well-established and widely used techniques for live cell isolation and single cell cloning—such as FACS, MACS, cloning by limited dilution, and so on—laser microdissection allows for capturing live cells and cell colonies without their detachment from the carrier. In other words, there is no need to prepare a single cell suspension before the isolation procedure using mechanical and enzymatic dissociation, which can affect cell fate after plating.
This feature of laser microdissection is desirable for stem cell research. We established a simple strategy for the efficient live cell isolation using the Leica Laser Microdissection platform. We were able to demonstrate not only colony formation from the isolated samples containing live cells, but also single cell cloning.
In this webinar, specimen preparation, laser adjustment, overall workflow, and limitations on live cell isolation by laser microdissection are discussed.
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View our on-demand webinar to learn about recent experiments using Laser Microdissection from Leica Microsystems
19th Oct, 2018

How to extract Image Information by Adaptive Deconvolution

LIGHTNING Image Information Extraction
Confocal Laser Scannning Microscopy (CLSM) is the standard for true 3D resolved fluorescence imaging. Fast optical sectioning using flexible scanning strategies in combination with simultaneous multi-colour, high sensitivity and low noise signal detection provides maximum resolution in the spatial and temporal domain. In combination with modern approaches to image information extraction this helps the researcher to mine as much information as possible from the images acquired. Image information extraction refers to intelligent procedures for image enhancement using a priori knowledge from the imaging system. From simple glare control and optical development to intelligent and ingenious model extraction, there are many ways to see more than just the image.
21st Sep, 2018

Increase efficiency in your cell culture workflow

Inverted microscopes for cell and tissue culture
Growing cells under lab conditions is the base for scientists working in the fields of cell biology, cancer research, developmental biology or any kind of life science and pharma research.
These easy-to-use microscopes allow you to configure an imaging solution for your needs with flexible condenser options and digital imaging documentation features.
When it comes to cell cultures there are a couple of differences between the solution´s instruments.
Use our cell culture wizzard, answer four quick questions and find out a solution that is just right for your lab!
Find your Cell Culture Solution
18th Sep, 2018

On-Demand Webinar on Fast FLIM - presented by Pr. Christian Eggeling

Super-resolved STED spectroscopy: New insights into molecular membrane dynamics
Speaker: Professor Christian Eggeling, University of Oxford, Molecular Immunology
Molecular interactions are key in cellular signalling. They are often ruled or rendered by the mobility of the involved molecules. We present different tools that are able to determine such mobility and potentially extract interaction dynamics. Specifically, the direct and non-invasive observation of the interactions in the living cell is often impeded by principle limitations of conventional far-field optical microscopes, for example with respect to limited spatio-temporal resolution. We depict how novel details of molecular membrane dynamics can be obtained by using advanced microscopy approaches such as the combination of super-resolution STED microscopy with fluorescence correlation spectroscopy (STED-FCS). We highlight how STED-FCS can reveal novel aspects of membrane bioactivity such as of the existence and function of potential lipid rafts, and how the new SP8 FALCON technology eases such measurements.
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View our on-demand webinar to learn about recent experiments using Confocal Microscopy from Leica Microsystems.
18th Sep, 2018

On-Demand Webinar on Fast FLIM - presented by Pr. Kees Jalink

Precise and lightning-fast spectral Fluorescence Lifetime Imaging at video rate integrated in a high-end confocal microscope
Speaker: Professor Kees Jalink, Netherlands Cancer Institute, Amsterdam
Förster (Fluorescence) Resonance Energy Transfer (FRET) has become a powerful tool to study protein-protein interactions and signal transduction in living cells and Fluorescence Lifetime Imaging (FLIM) is a robust inherently quantitative microscopy technique that can used to get reliable FRET data. In confocal microscopy, FLIM is typically read out by Time-Correlated Single Photon Counting (TCSPC). This requires expensive add-on hardware and is very slow.
In this webinar Prof Kees Jalink will present his research results acquired with the Leica SP8 FALCON, the first truly integrated solution for FLIM, capable of delivering benchmark results at least 10 times faster than conventional systems.
Says Prof. Jalink about his findings: ‘’We scrutinized the performance of the SP8 FALCON and report how it performed when recording agonist-induced changes in concentration of second messengers such as cAMP and Ca2+ with sub-micrometer precision at high speed allowing detection of even very fast signaling events. We concluded that the SP8 FALCON is ideal for fast FLIM screening applications in both fixed- and live-cell formats’’.
View Webinar
View our on-demand webinar to learn about recent experiments using Confocal Microscopy from Leica Microsystems.
28th Aug, 2018

Which Sensor is the Best for Confocal Imaging?

The Hybrid Photodetectors (HyD) are! Why that is the case is explained in this short article.
Biological research is concerned with life and, therefore, living samples, if at all possible, should be studied with microscopy. The fluorescent staining should be as sparse as possible to minimize any potential manipulative effect on the biological functions and parameters. The background should be pitch-black (given a clean sample) and the signal free of unnecessary noise. Therefore, confocal fluorescence imaging requires highly sensitive and photon-efficient equipment. Besides the optical lenses and filters, the sensor plays an important role in reaching this target. Good results were achieved so far with the somewhat moss-overgrown vacuum technology, represented here by PMTs (photomultiplier tubes), and more contemporary semiconductor technology, represented by APDs (avalanche photodiodes).
The Hybrid Detector (HyD) is a chimera of both technologies. It dexterously combines these two concepts and maintains the beneficial features of both:
short and equal pulses
extremely low noise, and
a large dynamic range
31st Jul, 2018

Microscope Resolution: Concepts, Factors and Calculation

Airy Discs, Abbe’s Diffraction Limit and the Rayleigh Criterion
In microscopy, the term ‘resolution’ is used to describe the ability of a microscope to distinguish detail. In other words, this is the minimum distance at which two distinct points of a specimen can still be seen - either by the observer or the microscope camera - as separate entities.
The resolution of a microscope is intrinsically linked to the numerical aperture (NA) of the optical components as well as the wavelength of light which is used to examine a specimen. In addition, we have to consider the limit of diffraction which was first described in 1873 by Ernst Abbe.
This article covers some of the history behind these concepts as well as explaining each using relatively simple terminology.
17th Jul, 2018

What is Photomanipulation?

The new Scientific Tool Box. Why is it important and how can you use it?
The term photomanipulation describes a wide range of techniques that enable the microscopist the transition from passive observer to instigator of events by offering a way of interacting with their sample via targeted illumination. Typically researchers are trying to observe specific processes of interest in order to understand the underlying biological process. Microscopists are often forced to hunt through large populations of cells or acquire hours of time laps footage before they’re able to observe events of interest and in many cases it’s simply not possible to observe certain processes using conventional microscopy techniques alone. Photomanipulation tools enable the microscopist to initiate biological events, precisely adjusting sample labeling, biological activity, local chemical environments and in some instances physically destroy parts of their specimen.
These capabilities are hugely valuable when it comes to unpicking complex, compartmentalized biological mechanisms. Photomanipulation tools enable the users to design advanced experiments to access additional layers of information from their specimen that are not possible from observation alone.
During fluorescence recovery after photobleaching (FRAP) the fluorescence tag of a protein is bleached by powerful excitation inside a region of interest (ROI). Subsequent resettlement gives insight e.g. into the turnover of the structure of interest. For fluorescence loss in photobleaching (FLIP) a ROI within a cell is bleached repeatedly. The bleached molecules spread out and the loss of fluorescence indicates, e.g. whether cell organelles are physically connected. During inverse FRAP (iFRAP) the cell region outside the ROI is bleached. Subsequently the non-bleached molecules spread out and can be monitored.
4th Jul, 2018

How to Choose the Right Confocal Microscope for Your Lab?

Free Buyer’s Guide: Confocal Microscopes for Life Science Discovery
Confocal microscopy has come a very long way since its invention more than a half-century ago. Today, with novel technology driven by leading imaging companies, it has become the standard for fluorescence microscopy. Choosing the right confocal microscope for your specific research requires the appropriate mix of features related to resolution, sensitivity, and speed. Confocal microscopy offers life science researchers a crisp, clear view into the inner workings of cells, structures, and tissues – going well beyond what can be seen through conventional optical microscopy.
In this free 16-page confocal buyer's guide, you’ll learn the key considerations for each step of the purchasing process, from budgeting to comparing different technologies to asking the right questions of sales representatives.
Bringing focus to your decision-making process so you make the best choice for you and your lab.
29th Jun, 2018

Introduction to Digital Camera Technology

Basic principles, properties, possibilities, and compromises
A majority of modern optical microscopy techniques require the use of a digital camera. By working with digital devices, researchers can observe specimens on a screen in real time, or acquire and store images and quantifiable data. A huge range of applications from basic brightfield imaging through to advanced super-resolution techniques all require cameras.
The performance and variety of available digital camera technologies has increased considerably, offering researchers a broad spectrum of detectors to address their scientific needs. The choice in digital imaging sensors can have a substantial impact on image characteristics, so it is important to have a basic understanding of how they work and differ from one another. Here we introduce the basic principles behind digital camera technologies commonly encountered in scientific imaging.
29th Jun, 2018

Eyepieces, Objectives and Optical Aberrations

For most microscope applications, there are generally only two sets of optics which are adjusted by the user, namely, the objectives and the eyepieces. Of course, this is assuming that the microscope is already corrected for Koehler Illumination during which the condenser and diaphragms are adjusted.
This article covers the components of the eyepieces and how to adjust them correctly to suit your eyes. Moving on to the objectives, we will examine optical aberrations and the four most common objectives which are corrected to overcome these anomalies.
Eyepieces and objectives are designed by microscope manufacturers to work in combination and optically complement each other. This is something which should be remembered if, for any reason, you are changing eyepieces or objectives between microscopes. The objectives and eyepieces of a microscope must work harmoniously with each other for optimal specimen imaging. When buying a complete microscope, the optical components will be designed and matched to complement each other to offer optimal viewing conditions for the user. Alternatively, if you are assembling a custom research grade microscope, then the choices of objectives on offer will determine which eyepieces are suitable for the range of objectives and vice versa.
11th Jun, 2018

Introduction to Widefield Microscopy

One of the most basic microscopy techniques is known as ‘Widefield Microscopy’. It is fundamentally any technique in which the entire specimen of interest is exposed to the light source with the resulting image being viewed either by the observer or a camera (which can also be attached to a computer monitor).
This article will set out to explain the differences between widefield (WF) and confocal microscopy looking especially at the differences between the imaging and illumination between the two systems.
Figure 1: Confocal vs. Widefield illumination. Confocal: Light from the light-source (Ls) is focused through a pinhole for illumination (Pi) and subsequently into the sample (S) resulting in a relatively small volume. Widefield: The whole specimen volume is exposed to light.
Microscope configurations for WF imaging will also be discussed looking at light paths involved as well as the problems of out-of-focus light. The more advanced WF techniques such as WF Super-Resolution will be considered below.
17th Feb, 2017

How Digital Microscopy can Contribute to Efficient Workflows for Microelectronics and Electronics

This report explains how users can benefit from cost-effectiveness over entire workflows with the digital microscope portfolio of Leica Microsystems.
Download this FREE report to learn about:
Research and development (R&D)
Product innovation
Process engineering, production, quality control and assurance (QC/QA)
Failure analysis (FA)
23rd Jan, 2017

Shed new light on your sample with digital microscopes

To better visualize product or product component flaws, different types of lighting contrast are exploited with optical microscopy.
From metals, polymers, to glass and geological samples, the need to efficiently discover flaws and defects can be critical in material sciences, as well as in production and quality control.
Download the free report to learn:
How recent developments in digital microscopy have led to a more efficient way of using contrast methods for inspection and testing purposes.
Various materials are used to show that if the feature of interest is seen more easily by enhancing its appearance with different types of illumination and contrast, then less time is needed for inspection and testing.
Example applications with integrated ring light or coaxial illumination system.
14th Oct, 2016

Fast and Reliable Inspection of Printed Circuit Boards with Digital Microscopy

What are the advantages of using a digital microscope for inspection?
In this report, we explain how a digital microscope makes the inspection workflow efficient, how you can view PCB components from different perspectives using tilting and stage rotation and how you can apply versatile illumination for multiple contrast methods and high dynamic range (HDR) imaging.
Download the free report to learn:
Rapid, easy tilting and rotation to view the sample from different perspectives
Integrated illumination for versatile contrast
High dynamic range (HDR) imaging
Large mosaic overview of the sample