Question
Asked 10th Jul, 2015
  • Tropical Agriculture Consultancy Services

Can someone give me information about the native/migratory duck species of waterfowl in Trinidad?

Species of waterfowl in Trinidad
Migratory patterns
Feeding and Nesting Habitats

All Answers (3)

13th Jul, 2015
Riyadh Mohammed
Tropical Agriculture Consultancy Services
Thank you sir, i will try them out. 

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Immunocytochemistry: Technique Questions
Discussion
7 replies
  • Anthony UndisclosedAnthony Undisclosed
Good morning everyone (at least for me)!
Some questions for the resident ICC experts (and knowledgeable beginners!) out there- as my latest dabbles in immunocytochemistry have been disappointingly unfruitful. I have much more experience with immunohistochemistry, but unfortunately there are some snags I'm experiencing in translating my IHC experience to ICC.
1. When washing your chamberslides/coverslips do you apply the wash buffer "indirectly" onto the slide/coverslip itself (ie. using a chamberslide or coverslip and applying wash buffer to the corner of the dish/chamber to best not peel off cells) or do you immerse the whole slide/coverslip into a chamber/bowl/liquid-receptacle with the wash and then let it soak (akin to traditional IHC)? This can also sort-of apply to the initial fixation as well- do you dip the slides into a receptacle containing the fixative or do you apply the liquid directly onto the cells/chamber?
2. When using chamberslides do you keep the chambers on during staining or do you remove them before staining? If your slides don't have hydrophobic barriers then allowing all the slides to sit in the same antibody 'bath' could help with ensuring consistent staining- but it comes at the cost of losing the flexibility of being able to have multiple conditions (like no primary antibody negative controls) on the same slide. Lately i've been concerned that some of my chambers "leak" as I notice some chamber wells have less liquid in them following incubation than others- leading me to be paranoid there is not just leaking but contamination of one chamber to another.
3. What confluency do you typically wait for before progressing with ICC? I am currently doing an experiment on fibroblasts and I'm at a loss for what percentage I should let the cells grow for. I don't want them overly confluent, but I'm also concerned that if they're not 'confluent enough' they may not have good adherence to their slide.
4. How do you remove your liquid from your chamberslides? Do you turn the chamberslide upside-down and (gently!) shake the liquid out into a sink, or do you aspirate the liquid out every time? When working with secure tissue you can use all manner of roughness when immersing and shaking liquid off slides- but with cells I'm scared of them falling off due to their delicate nature.
5. Are there any common reagents used in IHC/ICC that you would *not* use for ICC? Triton is very commonly used in IHC/ICC but many places say that it can be too rough at times- could this roughness translate to 'scrubbing' cells off the slide?
I anxiously await the input that any professionals or beginners (like myself) may have and are willing to share. Advice, comments, tips, tricks, suggestions, criticisms, and thoughts of any kind are welcome and greatly appreciated! Likewise if anyone has any questions of their own I encourage them to share and contribute.
I promise to respond as soon as I am able to any response that comes in.
Thanks! Your help is immensely appreciated!!!
Anthony

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