Cytology - Science method

Yes as mentioned in most of the replies above.

Please see details in some scientific papers such as http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3296804/

--A gastric cancer patient with positive cytology would be stage IV. -------

I suggest link for said query, might be useful for you.

1. https://www.researchgate.net/publication/237339713_Comparison_of_Common_Cytotypes_of_Andropogon_Gerardii_Andropogoneae_Poaceae

2. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3272458/

3. https://www.researchgate.net/publication/270965337_Environmental_correlates_of_cytotype_distribution_in_Andropogon_gerardii_Poaceae

4. https://www.researchgate.net/publication/259442688_Allium_oleraceum_in_Slovakia_Cytotype_distribution_and_ecology?ev=srch_pub&_sg=RBMCaSXQ0Q_Zm_fxc8VfZiy37LJLwVBTAuhjRc-RoQ9oYLR4cyyN-fYm929m87W1.p9uYQQsGc0oNBG9x23_RQjiURtCcG2O-9odvYp4nLPv_G4_Q7artbo1ZTJgm3QAP.2jnhmPPOlYqoxLeZ6Ruh92y9sSbumNg9EczJflQRrKVEfOmxlh5z3rkK5oOJSAt5

Regards

Abhishek Raj

Hello. We do not routinely take this exam. You've already seen this study. Maybe the article can help you.

Vet Ophthalmol. 2005 Nov-Dec; 8 (6): 401-5.

Conjunctival impression cytology in dogs. Bolzan AA.

In which species will you do the cytology ?

Good luck!

in 6s

Anyone who can host me in their lab to learn ICP measurement protocol? - ResearchGate. Available from: https://www.researchgate.net/post/Anyone_who_can_host_me_in_their_lab_to_learn_ICP_measurement_protocol#58c2bd40cbd5c278170e96c3 [accessed Mar 10, 2017].

Hi, 

    It's not mandatory to use FcR blocking reagents while staining with fluorophore conjugated antibodies. You will find lot's of literature supporting this. However there is a chance of non specific binding of antibodies to FcR on monocyte/macrophage, if the antibody, which you will be using is human origin (very less likely), then I believe you should consider use of FcR blocking reagents. If it is from other species eg mouse/rabbit (more likely), then there will be no point of using this. 3% BEA will be fine. 

good luck. Let me know if I can help with this further. 

I work with nanos in fish rather than seeds, but I saw that no one had replied to your question so I thought I would send a few suggestions to you. I have seen several talks on seed germination and growth with nanoparticle exposure and both of the methods you listed have been done (see publications below). The best method may be dependent on the properties of your nanoparticles (type, size, coating, etc.), species of interest (water plant, terrestrial, etc.), and environmental relevance (how would your species encounter the nanos in the environment?).

Good luck!

- Melissa

Dear Ivan,

I have already found somebody who will co-operate with us. I want to test a human Pneumocystis antibody solution by IHC. Aim is to use this solution for immuno-precipitation for Pneumocystis in pig BALF. The initial steps are the establishment of a functional IHC on human samples and on pig samples.

If I need the mouse sample, I come back to you. Thank you for offering this.

I have passed on your greetings.

Best regards,

Christiane

If cytospin after fixation is not possible:

You can do IF-taining in tube (1.Antibody, washing steps 2.Antibody, DNA-staining ...)

and fix the cell before microscopy on the slide with low melt 0.5%-1% agarose.

Hi, Nader,

Cytogenetic plant (wheat) techniques, namely, Fixation, Pretreatment, and Staining, please, see at site

Godspeed

climate change effect the physiology of plant

You can make a cell block the technique you can found everywhere. 

But in my experience, instead of using commercial gel (ex Histogel), we use Agar gel 3%, it works well. 

these cells are the size of lymphocytes with very dense nuclei, I think  they could be apoptotic cells, perhaps due to therapy?

you can try confirm apoptotic cells by immunocytochemical detection of c-caspase 3?

check this article http://www.thelancet.com/journals/lanres/article/PIIS2213-2600(14)70050-5/fulltext?rss=yes , where similar cells are shown

Thanks for your question - we're not performing any screening for this study, just interviewing women and HCPs who perform testing about their experiences of the test. (The project is in the UK, so the tests the women will have experienced will be liquid based cytology, proceeding to an HPV test only when abnormal cells have been detected.)

Dear Qian,

That is a common problem with acinar structures of pancreas and salivary glands which makes immunostaining so difficult. I have worked on both and what helped me was using a prolonged incubation in serum-free protein block (I have used Dako) and background-diminishing antigen diluent from the same source. I am sure that there are similar preparations from other manufacturers, but for pancreas I have only used those two. You may have to do quite a few experiments to establish the working protocol for timing of incubations with protein block and concentration of your prime antibody (after blocking and using this diluent - usually higher than you would use for another type of tissue).  

I have attached some useful papers. Go through it.

Regards

Abhishek Raj

Telomeres are the very end of linear chromosomes and can be defined by sequence (in most organisms T-rich on the "sense" strand), these are covered by specific protein complexes such as Rap1-Rif1-Rif2 and Cdc13 (POT1 in humans). This sequence can be extended by telomerase (telomerase also contains an RNA which is complementary to the telomere sequence which helps to recognize telomeres and extend them). Now, subtelomeres are less well defined on a sequence level, it's mostly the region next to telomeres, these can be repetitive in nature and they are often heterochromatinized. Hope this helps!

Sorry I thought you wanted to analyse immune cells in blood. In that case 

Try adding some dextran 40 to your media at about 30mg/L. Heparin can cause RBC aggregation so don't use it in the media that you use in the microfluidic device. You can wash your RBCs in PBS twice and resuspend in PBS with dextran 40. You could also try increasing the shear in the device as well (higher flow rate or smaller dimension in part of the fluid path) to disaggregate the cells. RBCs often aggregate in low shear environments.

Skin lesion, lump, or ulcer that do not resolve in 14 days located:

On the tongue, lip, or other mouth areas

Usually small

Most often pale colored, may be dark or discolored

Early sign may be a white patch (leukoplakia) or a red patch (erythroplakia) on the soft tissues of the mouth

Usually painless initially

May develop a burning sensation or pain when the tumor is advanced

Behind the wisdom tooth

Even behind the ear

Additional symptoms that may be associated with this disease:

Tongue problems (moving it)

Swallowing difficulty

Mouth sores

Pain and paraesthesia are late symptoms.

Hello Inaki,

in case you don't get your chambers, an alternative is to mark lines on the bottom of your petri dish, e.g. in 0.5 cm distance, can be by a fine pen, or with a scalpell. Or to buy transparent thin foils with such patterns printed, or prepare such foils yourselfe.

When upright illumination, we just print patterns on paper (usually black background + white lines, but can also be opposite) and put that unterneith of petri dish, for counting. Just in case,

Arthur

Dear Murali, thank you.

This is a good addition to my question.

Dear @Claudia Mazo.

Thanks for your reply.

Vyacheslav.

sorry I am not a botany is not my speciality K.

What do you refer with "large": battery, capsule or tubule? The largest undischarged nematocyst capsule I ever measured is ca. 55 x 9 µm, which is large for a benthic hydroid, but not quite impressive compared with nematocyst capsules found in some Siphonophores [e.g. Purcell (1984) reported heteronemes capsules of ca. 150 µm] 

Kitatani et al (2015) reported tubules of several mm in some Carybdeid species. Have you checked it?

 

Kindly elaborate about the essence of your project. Are your findings against the soul theory mentioned in Geeta or Kriya Yoga techniqs....

I still feel as and when Ovarian lesions are excised, one can prepare a set of good well fixed slides for study and learning from classical lesions.

Dear Mehmet,

Because of the nature of HPV (epitheliotrophic and local infection), you can find the HPV DNA from direct sampling of tumour tissue by PCR assay. The stability of DNA let you detect the HPV DNA from frozen (at -20 degree) or paraffined sample.

OBA TOYIN MUSTAPHA

Department of Plant Biology, Faculty of Life Sciences, University of Ilorin, P. M. B. 1515, Ilorin, Kwara State, Nigeria

The author can be requested for the full form of AJMNS

TENT buffer is used to extract DNA, the composition should be:

10 mM Tris-HCl, 0.1 M NaCl, 1 mM EDTA, 5% [v/v] Triton X100, pH 8.0

Please have a look at this useful ResearchGate link.

Hi there,

Alkaloïds are secondary metabolites mainly produced in plants. As these molecules don't participate to primary metabolism, they circulate freely into the whole organism. Their role is purely protective against insects and mamals due to their toxicity on non plant specific targets.

The GOI Operational framework recommends VIA for cervical cancer screening till the time an appropriately priced and suitable HPV test becomes available.

Yes, for Bonferroni correction you did correct. Basically, here are 2 ways of doing it and both lead to the same result: one way, as you did: when deviding threshold level (0.05) by number of tests. And the second way: to multiply obtained p-values for each test by the number of tests and if the adjusted p-value is still below 0.05, consider as significant. This is about Bonferroni correction.

But now, before doing the Bonferroni correction, clarify for yourself: did you do multiple logistic regression of univariate? If you did multiple logistic regression, you do not need any further correction. As I understood, you combined them, meaning that you did multivariate logistic regressoin analysis. So, just use the p-value you obtained. You could apply the Bonferroni correction to the initial analysis, when all biomarker were analysed between normal and cancer serum.

Dear Dung

In the attached papers, apomixis frequency and ploidy levels in Poa and Centotheca using flow cytometric techniques were examined, which I hope will be of interest to you.

Dear Dr. Roughani

In addition to herbaceous plants, this problem is also present in some fruit trees, and the cell wall is not broken down by the enzymes. In the article I sent, they present a new enzyme digestion protocol that is fast, efficient and flexible.

Use epigenetic modulator both DNMT and HDAC like 5-Aza cytidine (500nM) and SAHA (500nM to 2uM) for 24 to 48 hours in combination.

Hello Hamdia,

Saccomanno's Fixative is an excellent pre-fixative for cytology specimens and the fixative of choice for collecting and fixing sputum specimens for cytology. This fixative in in fact, one of the most widely used fixatives in cytology that is recommended for cytology specimens such as sputums, urine, FNA'S, bronchial washings, pleural and peritoneal fluids, and ThinPrep™ preparations. The formula is accessable at the website indicated below:

Saccomanno's fixative is 50% ethyl alcohol which contains approximately 2% of Carbowax 1540 (Union Carbide Corporation, UCAR). Carbowax 1540 is solid at room temperature, with a melting point of 43 to 46 C. To avoid having to melt it whenever the fixative is prepared, a stock solution can be prepared by melting Carbowax (melted in an incubator or hot air oven at 50 to 100 C) and adding it to an equal volume of water or 50% ethyl alcohol. The mixture will not solidify. Saccomanno's fixative can then be prepared with 430 mL of water, 530 mL of 95% ethanol, and 40 mL of the stock Carbowax solution. Some light green SF or fast green FCF can be added to color the fixative. Koss warns that the denaturants in reagent alcohol may cause excessive hardening of mucus.

ref: Histology FAQ

Dr. John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada

With best regards,

Dept of Pathology

Faculty of Medicine and Surgery

University of Santo Tomas

Manila,Philippines

Simply, from callus appearance. Embryogenic callus white and granuular apperance while the other callus looking yellow and friable.

Have serum alpha-fetoprotein (AFP) and beta-human chorionic gonadotropin (HCG) levels been checked?

FNAs in most cases may help. However the size of the mass raises additional risk of torsion.

Laparotomy with biopsy is a gold standard to obtain a definitive diagnosis.

Interpretation of cytology is based on the complex of features, including cellularity, architecture, nuclear features, nucleo-cytoplasmic reatio, presence of mitosis and/or necrosis etc. You are right that identifiation of malignancy in some cases is based on specific features, typical for some cancers. In addition, you can use motelular testing when having indeterminate cytology

It may help:

You could use a silane such as 1H,1H,2H,2H-perfluorodecyltrichlorosilane, commonly abbreviated as PFDTS.

It is rather inexpensive and I think it would do the job.

I don't have an universal answer. If you have a touch it means you have both tissue sample and imprint taken from this sample i.e. both histological and cytological material whereas the FNA is cytology alone. However several FNA needle passes may be done in different directions so it's sensitivity could be higher than that of core biopsy, and the cell block could be (I would say should be) prepared as well. The question is whether your reference laboratory is able (both practically and legally) to perform the mutational analysis in the FNA material (cell blocks I mean) in the cases it is needed.

Sincerely,

Alexander Shtabsky

TinPrep: The technique is simpler but expensive. It is less cells then Surepath.

SurePath: More handwork but cheaper. More cells, no problem with bloody and mucoid specimen. You find more adeno ?

Dear Ahmed,

apologize if my reply was not quite revealing. I once had a similar problem for my semithin sections and the problem was that semary "dirt-particles" disturbed the viewing as well as imaging/documenting of stained sections. So I decided to do the same... returning ... and got another batch of slides which was ok. Nevertheless I always clean (HCl-& alcohol,washings in distilled water) slides to be used (even if they are taken from an originally packed box). This helps(ed) always not only for correct sticking of the resin sections but also in achieving good staining results.

Have a nice weekend,

best regards, Wolfgang

I suppose that we are considering extra-hepatic bile stricture.

I think that a preoperative cytological confirmation is not necessary. The incidence of benign biliary stricture in absence of previous surgery or calculi is less than 10%. The chance to have preoperative positive cytology is about 50%. In case of a report "negative for tumor cells" are we so confident in suggesting follow-up or stenting? We have to clearly inform the patient, what about: you have a 90% chance to have a malignant stricture.

It's extremely important to have a well done pre-op work up and than we can offer surgery without hesitations

There are many members of the genus and also Anaplasma in animals that infect a broad variety of blood cell types.

There are various biomarkers for inflammation in the GI tract. Calprotectins may be used as a biomarker for evaluating in GI inflammatory disorders.

Attached is an article that may be of some used to you when looking for markers of inflammatory conditions of the GI tract.

You don't happen to have the publication available somewhere ... and a photo of Thomas Ashworth?

I too, have wondered about this! If an abstract is published, in the proceeding of a research gathering, it definitely is worth some credit, in terms of points or else. But, I have not come across any such quantification.

A is possibly sticky chromosmes at Anaphase and not metaphase as the chromatids are already migrating to opposite poles. B. is an abnormal stage in prophase. For clearity and easy identification of the abnormal chromosomes i suggest that the magnification be increased. Thank you

logically, We can use barr body as marker to discriminate between male and female nuclei. histone macroH2A1 staining is used to identify barr body due to histone macroH2A1 content of histone protein which play important role in silencing of whole X chromosome.

Best regards.

In PSG tech, the TEM studies cost of per sample is 2800/- with tax.

Since the size of MVs and exosomes close (or under) the physicial resolution limit of visible light microsocpys, you have to mark the vesicules with fluorescent markers to make them visible - so you will not see directly the subcell. ves.-s, but their fluorescent halo. The fluorescent markers cen be specific (such as fluorescent protein conjugated specific antibodies), or aspecific (using lipid markers, such as Annexin V for phos. serin, or other PKH stains). With the first method you can detect specific population of subcell. ves.-s, while with the other you can detect all of them. It also important to make a control of specificity of your stain, such as using detergent (e.g. Tween, Saponin, etc.), to eradicate the vesicles - in this case you wont see fluorescent marks. A third approach can be using intravesicule markers, as dr. Ganglum wrote.

By the and, let me recomend a good protocol oriented article: Detection and isolation of cell-derived microparticles are compromised by protein complexes resulting from shared biophysical parametershttp://www.ncbi.nlm.nih.gov/pubmed/21041717

With best regs: Csaba Timar

I am dealing with invertebrate tissues but my supervisor ifor bowen has a lot of papers about cancers you can search for i.d. bowen and also cathrin saraf

You are correct that imaging software is more accurate. Try Image J, its free and works on many platforms

Sorry , Idont no

Hi,

ThermoElectron Corp Cytospin 4 CytoCentrifuge and its a good one.

Good luck.

What type of microbes u want to perform a cutinase assay?

For adherent cells I suggest to use chambered coverslips (http://ibidi.com/xtproducts/en/ibidi-Labware/Open-Slides-Dishes:-ibidi-Standard-Bottom/m-Slide-8-well) or porous membranes (https://de.vwr.com/app/Header?tmpl=/life_science/hts_transwell.htm).

hi Alexandra

sorry about delay. I have problem with site.

I don't no whole of your process. but i think you can use nylon wool for separation lymphocyte.

Besides the usually used Trypan Blue also can be used: methylene blue (absorptive stain), indigo carmine (contrast stain), Cresyl violet; Erythrosine (Red No 3), Eosin (both are exclusion dyes), Propidium iodide; 7-aminoactinomycin D, Alizarin Red, fluorescent dimethyltin(iv)-alizarin red S complex....all depending on the exptl. necessity you have. Even injections of lead acetate as an intravital stain of calcification sites have been described. Perhaps interesting to you also would be reading: Time-Saving Benefits of Intravital Staining

@ http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2901106/

Best wishes and regards,

Thanks David and Christoph

Tibor, just asking about your standard PAP-staining: which one do you use (when reading:

with 3 procedures: 

i)    Procedure 1 (Standard Method)

ii)   Procedure 2 (Modified Pap Procedure)    and 

iii)  Procedure 3 (Rapid Economic, Acetic Acid, Papanicolaou Stain Method)

I am wondering why you did not get any answer since 2012! (but admit that I found your question some minute ago....)....would recommend, to edit your question and add some keywords more: e.g.: Histology, Histochemistry, smears, Staining   to eventually increase readership.... best wishes and good luck...W.M.

NB: I haven't done PAP-staining during my former professional career...but perhaps I could help -at least knowing the protocol of your PAP-stain....

Hello Vinicius,

I'm working with coelomocytes of Paracentrotus lividus.

Christopher

Yes

The HPV virus has to reach the basal cells of the cervix. Therefore, there must be some form of break in the epithelium for the HPV to reach the basal cells. Cervicitis is one way in which there can be a break in the epithelium. A similar situation exists in throat cancer and the relationship between smoking and drinking spirits. The smoking and spirit drinking results in a break in the epithelium and therefore HPV can reach the basal cells. In cervical cancer it does not have to be cervicitis and the process of metaplasia is also thought to be associated.

Hi,

Maybe you should also take care of the quality/purity of your plasmid DNA, e. g.  by using an endotoxin-free plasmid kit when preparation from bacteria. The amount of DNA might also be a critical factor. Also, try out transfection without serum, if you did not so. Good luck.

Blood collection done with precautions for asceptic conditions will not get contaminated. It is better to use vacuuatainer, and if it is manual method i.e. collecting with syringe and needle and opening the collection vial, then we have to ensure that; there are no direct air currents like fan etc. nearby, use two burners, light them some 10 minutes before and open the vial between the two, use only Sodium Heparin as anti-coagulation factor, and most imp., mix it well and store at room temperature only, set the cultures not later than 36 to 38 hours, and it should yield proper metaphases. Also when you add the sample to culture media see if it is very old, in that case  L-Glutamine can be added to ensure active proliferation.

Please see these papers. They may help you out.

Cancer Letters vol.323(1) 2012 20-28.

Good Luck!

I agree with Laura and Shahzad, it is not easy to detect secondary spermatocytes, specially when tubular sections are not completely obliques (since maturation stages are mixed). Performing complementary histological sections with PAS staining  could help you to differentiate stage XII, and thus secondary spermatocytes. But tubules must be completely rounded and with the lumen visible. You can look up my last work in which I combine immunohistological techniches with PAS staining in mouse testis.

Try to fix with only 100% of methanol.  PFA (fixative) consist sodium and formic acid which usually causes some shrinkage and/or distortion of the specimen. This might caused the distortion of the nucleus. Most of the staining (e.g: Wright's staining) for thin blood smear (obtained using blood smear or cytospin) requires methanol as the fixative agent.

All the best.

cytogenetics :the study of chromosome and the related diseases caused by the abnormal no or structure of chromosome.

chromosomes can be identified by their size,shape and banding patterns such as Q,G,R and C-Banding.

Karyology:the study of the cell nucleus, organelles,structure and its functions.

Karyotype has the formula.such as 46,xy,....

The LBC was developed in an attempt to reduce gaps in conventional cytology promoting the use of cleaner slides, no superpositions of cells or other obscuring elements. This is due to the filter system where only epithelial cells are retained resulting in a monolayer or a thin layer slide.  In this cytology sampling is similar to the conventional one. However, in order to decrease the presence of artifacts, the cytological sample is dispersed in a liquid suspension that after centrifugation, passes through a filter. Cytology is obtained from the suspension and prepared in a monolayer with the morphological structure of the preserved cells. Note that the sample preparation was adequate and correct, if so, you need to differentiate artifacts of cells. The reading of the blade is about the same, the advantage is the precision with which the cytological blade shown. Depending on the site collection, it may be some artifacts that were released from the cells, or technical artifacts. But as for your question, the red cells may be cells that stain of non-specific way, try changing the observation area and see if it repeats the rest. It is difficult to say something when we do not have the vision of the blade.

Levan et al. 1964 is generally followed. 

As our DNA or any other kingdom's or species' DNA is made by same kind of chemistry i strongly believe the DNA repair capacity matters and have a great effect on genomic integrity. The strand breaks (single or double- if they are not repaired) of DNA can be visualized as Chromatid Breaks in the metaphase (by observing Chromosomes). There are lot of publications which can interpret the possibility of gaps and breaks in the chromosome arms (Chromatids) !

1) On the nature of visible chromosomal gaps and breaks. 2004) Savage JR. Cytogenet Genome Res. 104(1-4):46-55.

The above article is just an introduction, if your question concentrates on how the chromatid break happens .?

The damage to the human genome incurs on the order of 1, 000-1,000,000 DNA lesions/cell/day ( Molecular Cell Biology by Harvey Lodish, 4th edition 2000) Most of the lesions are caused by endogenous sources, including reactive oxygen and nitrogen species (ROS, RNS) that can oxidize cellular macromolecules including lipid, protein and nucleic acid where one or the other takes part in the integrity of genome and DNA damage signals

"in common man's words' ...for example..... what does affect the normal cell.?"

the lymphocytes can be affected by the air u breath in

Skin can be affected by the sun light u get exposed

squamous cells in the the passages of the respiratory and digestive tracts, and the linings of the hollow organs of the body can be affected by the HOT DRINKS you take and the SMOKE u inhale.. and they get repaired...this is all normal stories Check out the below article it might be useful

2) Schanz S , Flockerzi E, Schuberth K, Rübe CE (2014) Genetically-Defined DNA Repair Capacity Determines the Extent of DNA Damage Accumulation in Healthy Mouse Tissues after Very Low Doses of Ionizing Radiation. J Carcinog Mutagen 5:196. doi: 10.4172/2157-2518.1000196 good luck!

Thank you so much Mr. Silva and Emin for recommendation, I am going to download it

then I need your advice later

Thank you :)

The nature of chromosome pairing at meiosis in some tuberiferous Solanums (2n, 3n and 4n) is described. A number of abnormalities have been noted at meiosis in the diploids including the formation of multivalents and univalents. Although the occurrence of univalents as early as diakinesis suggested that chromosomal differences do exist, the evidence from the diploids as to the nature of the basic genome in Solanum is not clear cut. An examination of meiosis in triploid hybrids (4n × 2n) revealed a considerably higher value for the mean number of bivalents plus trivalents per cell at metaphase I (14.2; 13.5; and 13.3) than has heretofore been reported. The maximum number of such associations recorded was 16. It appears that in the hybrid between the induced tetraploidS. chacoense (n=24) andS. neohawkesii (n=12), at least 8 out of the 12 chromosomes from the diploid parent are capable of exhibiting partial homology. The analysis of pairing in the triple hybrids ((6n × 2n) × 4n) is more complicated. However, there are some indications that autosyndetic pairing within the complement of chromosomes derived from the 4n parent may exist. The occurrence of pairing within the haploid set of chromosomes derived from one diploid species appears to be suggestive of the possible existence of two subgenomes within the haploid genome but further studies of the whole genus are required for a verification of this possibility.

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@Please enlist the difficulties/observations you have come across.

Regards

We follow the same basic steps in reporting:

1- Adequacy: assessed by many alveolar macrophages (not columnar and certainly not oral squamous). Number is not absolute and depends on method of preparation, but ~50 per slide would be a minimum for me

2- Presence or absence of evidence of viral, fungal or bacterial infection, such as inclusions, inflammatory cells, frothy background of Pneumocystis. etc. Abundance of neutrophils, eosinophils is noted.

3- Presence or absence of Atypical or malignant cells and type.

4- Specific comments in case of possible aspiration, allergy or pulmonary hemorrhage (as outlined in the previous response)

The typing of lymphocytes as to T and B by immuno is not reliable. It should be done by Flow cytometry and needs a separate sample, specifically processed for that, This is unlike FNA, where the samples are much more cellular and can be preserved rapidly, thus preserving the integrity of surface markers.

(see Erozan: Pulmonary Cytopathology, just published by Springer, 2014)

The following article gives answer to the question

Sarcoma. Jun 2003; 7(2): 87–91.

doi: 10.1080/1357714031000081207PMCID: PMC2395518Malignant Glomus Tumour: A Case Report and Review of the Literature

Annarosaria De Chiara,1 Gaetano Apice,2 Stefano Mori,3 Giustino Silvestro,4 Simona N. Losito,1 Gerardo Botti,1 and Vito Ninfo5

1 Department of Pathology, Istituto dei Tumori di Napoli ‘G. Pascale’ di Napoli, Via M. Semmola, Napoli, 80131, Italy, 2 Division of Medical Oncology B, Istituto dei Tumori di Napoli ‘G. Pascale’ di Napoli, Napoli, Italy, 3 Division of Surgical Oncology B, Istituto dei Tumori di Napoli ‘G. Pascale’ di Napoli, Napoli, Italy, 4 Division of Radiotherapy, Istituto dei Tumori di Napoli ‘G. Pascale’ di Napoli, Napoli, Italy, 5 Department of Pathology, Facoltà di Medicina e Chirurgia di Padova, Padova, Italy, Annarosaria De Chiara, Email: moc.liamtoh@araihcedanna.Corresponding author.

Author information ► Copyright and License information ►

Copyright © 2003 Hindawi Publishing Corporation.This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

This article has been cited by other articles in PMC.

Abstract

Purpose: Glomus tumours are characteristically benign solitary tumours. At our knowledge, about 23 reports are present in literature regarding the malignant counterpart, but only a minority developed metastases. We describe a locally aggressive glomus tumour with lymphnode metastasis.

Patient: The patient was a 40 year-old man presenting a 1.5-cm lesion on the right wrist incompletely excised and a recurrent tumour, 4 × 2 cm in size, removed after 9 months, for which he received radiotherapy. After 2 years he developed an axillary lymphnode metastasis.

Results: Histologically, both tumours (primary and metastasis) were similar. There were sheets and nests of uniform small cells with scant eosinophilic cytoplasm and round to polygonal nuclei; there was some degree of pleomorphism and the mitotic index was high (up to 18 m/10 HPF). The tumour cells were positive for vimentin and smooth muscle actin, but negative for desmin, NSE, Factor VIII, chromogranin, cytokeratin. Remarkably, in the primary, the cells strongly expressed p53 (70%) and MIB-1 (35%).

Discussions: In many reported malignant cases, the histology of the tumour cells suggested that they were malignant, yet the clinical course has been benign. Carefully reviewing the literature, it seems that actually we have enough histological criteria to identify the cases with biological adverse outcome. Those unfortunate cases behave as high grade sarcomas and therefore may deserve an aggressive therapeutic treatment.

In theory, you might be able to identify your acanthocyte population simply by using Forward and Side scatter, as these cells appear to be smaller and possibly more granular than typical erythrocytes. Have you ever looked at your control and toxin treated cells via SSC/FSC? Happy to have a look, if you could post a picture of your flow cytometry data.

AnnexinV is used to detect phophatidylserine exposure - this is for example the case for apoptotic neutrophils that can be thus phagocytosed by macrophages. I have found one article, where AnnexinV was used to detect changes in the phospholipid structure of erythrocyte membranes (PMID: 8562945), so might indeed be worth using this as a stain.

Could you post some example images?

Prepare always fresh specimen- Peripheral smears  from a freshly  drawn blood specimen (EDTA purple top tubes). Allow smears to air dry completely before staining.Please mix the Wright Stain and buffer mixture before blood smears are

made or before the smears are air drying. Combine 15ml of Wright Stain with 75ml of Wright Stain Buffer, mix well and allow to stand for at least 10 minutes. Make sure that a metallic sheen is observed on the surface before staining, Place freshly prepared and air dried blood smears in a manual  staining rack -->Place slide rack with air dried blood smears in Methanol for 30  seconds---> (Adjust timer for 30 seconds--> Take a disposable pipette and flood the Wright Stain on the

appropriately labeled slides. Set timer for 3 minutes. Place oxidizing Wright Stain and Wright Stain Buffer Mixture on Wright stained slides laying on slide rack (Displacing the Wright .Stain off the slides with the pipette filled with Wright Stain/buffer mixture and viewing a metallic sheen on the top of slides.Set timer for 6 minutes. Place slides in Wright Stain Buffer for 1.5 minutes. Set timer for 1.5 minutes. Retrieve slide rack with stained slides from buffer rinse and allow to air dry before examination with immersion oil and 100X objective.Wipe excess stain from back of slides with  methanol soaked gauze, being careful not to wipe off the stained side of the

slides.Place on slide envelope or slide flat/folder and proceed to the microscope room. Place 1 drop of immersion oil onto slide to be viewed making sure

it is completely air dried after staining. 

What do you mean by the cells are broken?

 

 

Hi Edelmiro,

I'm not a chemistry, but i think that you could add to your compound (and as control even to the others drugs that don't work) some specific sequence for a mitochondrial matrix enzyme. Then you could verify that only your drug is it inside the matrix because it will be the only that will give you the specific reaction. In this way you will demonstrate that only your compound is inside the matrix and not the other. Probably is not easy to do, but you could try. On the other hand, you wrote that your compound block an enzyme inside the matrix. You could evaluate some substrate of this enzyme and you should find a different amount ( phosphorylation- cleavage - i don't know what is your enzyme )compared to the untreated or the treated with the un-functional drugs.

i don't know if they are good ideas, but with so few informations i can't help more.

regards.

Francesco

Instead of acetocarmine, you can go with "Feulgen staining" which stain only nucleic acid.

Thank you Danilo. I noticed this artefact on CK7 staining as well. Might be an effect CK7 has on certain cultured cancer cells.

Dear James

This article enclosed may be useful for you

Quantitative analysis of fine needle aspiration biopsy samples

The genus Crocus L. belongs to the large family Iridaceae and is a systematically problematic genus. In the Old World, about 100 species are known (Harpke et al., 2013). Although molecular studies have been increasingly used to examine the phylogeny of living organisms, they have only recently been applied to the genus Crocus (Petersen et al., 2008). DNA markers have been used to characterize germplasm collections, inform breeding programs, and facilitate genetic diversity studies and taxonomic analysis. The utilized methods include interretrotransposon amplified polymorphism (IRAP) (AlaviKia et al., 2008), random amplified polymorphic DNA (RAPD) (Grilli Caiola et al., 2004; Beiki et al., 2010; Rubio-Moraga et al., 2010), amplified fragment length polymorphism (AFLP) (Zubor et al., 2004; Erol et al., 2011; Nazzal et al., 2011), intersimple sequence repeat (ISSR) (Rubio-Moraga et al., 2010), and simple sequence repeat (SSR) (Rubio-Moraga et al., 2010; Nemati et al., 2012) analyses. Among these molecular markers, AFLPs were found to show high levels of polymorphism per primer pair and yield high resolution and reproducibility (Meudt and Clarke, 2007). Fore more information follow this link: