Questions tagged [staining]

Set of methods to visualize certain properties (structure, gene expression, potential changes, etc.) of organisms or single cells.

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40 views

How a pathologist would analyse this H&E image?

I am working on a project which involves writing computer software to analyse histological images. A typical image looks like this: It is a Hematoxylin and Eosin stained biopsy of breast cancer ...
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DRAQ5 and PI incompatible?

I'm interested to analyse hemolymph cells from a marine mollusc. I wanna count only "alive" cells so my strategy was the following: DRAQ5+ (nucleated cells) and PI- (alive cells) and identify ...
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Is there any double staining method to visualise separate colour for plant tissue and fungal tissue?

Is there any double staining method for developing separate colour on fungal tissue and plant tissue when the said 2 types of tissues are intermingled? Unlike my previous question, this question not ...
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How can I stain only the Chitin but not Cellulose or Glucans?

Is there any stain, that binds specifically to chitin but not to cellulose? (for differential staining and double staining purpose) It seems the lactophenol-cotton blue, aniline blue, trypan blue etc ...
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Re-stain slice using a different secondary antibody?

I stained a free-floating ~300uM brian slice using 4 different 1º antibodies (rabbit, chick, mouse, rat). For the mouse primary, I mistakenly used an Alexa-350 secondary, when I should have used 594. ...
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1answer
379 views

Flow Cytometry Channels

I did a flow cytometry experiment for the first time. I had three conditions: (1) unstained healthy monocytes (2) healthy monocytes stained with green/red (488/570) from the Live/Dead thermofisher kit ...
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82 views

Dark and bright sides in fluorescence microscopy images?

I have used Phalloidin to label F-actin in ethanol fixed cells ( I know formaldehyde might have been a better choice for fixation). I am new to new to this and I am experiencing that one part of image ...
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288 views

Common stains for fungal microscopy

I recently got a light microscope to start exploring microscopic features of fungi (and to explore microscopy in general). What are some of the most common and easily available stains used when ...
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1answer
7k views

Cyanobacteria: Gram negative or Gram positive?

My (school) textbook states that cyanobacteria ("blue-green algae") are Gram positive (no references provided). They mention this at several points over the chapter we're doing so it doesn't look like ...
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1answer
759 views

Flow cytometry data Analysis [closed]

everyone. Since I am new in the flow cytometry technique, I am not really sure how to analyze the result. We use cell U-937 Lymphoblast.All Cells Could Anyone help to explain me this one? Thanks ...
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1answer
395 views

Quantifying DNA in a band on gel electrophoresis

I am a first year science undergraduate and I am just asking this as a more theoretical question rather than for carrying out a protocol, so I would appreciate if answers do not involve lots of ...
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1answer
17k views

Are acid-fast bacteria Gram-positive or Gram-negative?

Both procedures involve using a stain, decolorising agent, and then a second stain. I am tempted to conclude that acid-fast bacteria, which resist decolorisation (eg. by strong carbol fuchsin), also ...
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Is it possible to color sperm cells so they look more clear under a microscope?

Im working on something that requires me to check the velocity of a sperm cell under a microscope but I imagine it would be very hard for a program to keep track of the sperm cells.
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295 views

DAPI staining showing unknown artefacts?

Dear fellow researchers, I have been facing some problem with staining nucleus with DAPI. I use the standard protocol for fixation 4%PFA+0.1% TX-100+2%BSA. I use 0.2ug/ml conc. of DAPI for 5 minutes....
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55 views

How does gold chloride stain neuromuscular junctions?

Gold chloride is used in muscle biopsies to stain the NMJ. What is the substrate that it binds to?
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1answer
2k views

Forgot to vortex antibody before staining

Ugh. Did an immunofluorescence experiment last weekend, forgot to vortex both my primary and my secondary antibody solutions. And my final result looks dimmer than it should be. Is it possible that ...
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1answer
3k views

Help analyzing SDS-Page gel

In this experiment, we transformed a truncation of the NFAT protein sequence into a plasmid vector to be expressed in E.Coli as a fusion protein with GST. We also attempted to transform the normal ...
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1answer
92 views

Genomic instability of cancer cells in h&e image

Is there are any books or papers on the topic of visual differences(analyzing H&E microscopy) of high grade cancer cells vs low grade cancer cells vs non-cancer cells? In particular, I am trying ...
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83 views

How to use DRAQ5 and MitoTracker CMXros dyes to stain turtle erythrocytes?

I am working on turtle species and want to use DRAQ5 and MitoTracker CMXRos to stain turtle nucleated erythrocytes. Can anyone suggest me a protocol to carry this stain out? Would the two stains ...
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1answer
2k views

Staining cells for FACS at 4 degrees or ambient temperature

I'm sorting a 293 derived line. One thing that is worrisome for me is cell viability after the sort. Usually I have been staining and washing at room temperature (on the benchtop) on a nutator. I have ...
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1answer
494 views

Choosing the best assay for my experiment? [closed]

Which of the following assays can be used to determine whether cell death specifically due to apoptosis has occurred in a given cell sample? Choose all that apply. WST-1 Assay Caspase Colorimetric ...
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1answer
757 views

How does the structure of the pancreatic acinar cell relates to its function?

So the pancreatic acinar cell synthesizes, stores and secretes digestive enzyme precursors called zymogens e.g. pepsinogen. The structure of the acinar cell shows that there is an apical and basal ...
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2answers
2k views

Barr body Giemsa staining

Not all the female cheek cells show Barr Bodies when stained with Giemsa stain? Only 30 to 40% female cells show Barr Bodies. Why? Our Experiment: We washed mouth twice with listerine to reduce ...
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2answers
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Is there a stem cell dye for education purposes?

I need a dye that will stain stem cells (for this purpose the stem cells will proquired from chicken bone marrow). This will be an educational hands on activity for young teenagers so ideally the dye ...
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1answer
2k views

Does methanol fixation deform cultured cells?

I use methanol fixation (@ -20⁰C for 10 minutes) when performing immunofluorescence assays on cultured cells. Generally speaking, this results in very good antibody staining. However, the cells tend ...
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2k views

Hoechst red and blue spectra - what does the red emission tell us?

I understand that the blue spectrum is emitted when Hoechst 33342 binds to nuclear DNA. But what exactly is the red spectrum telling us? I read that the red spectrum can be indicative of apoptosis, ...
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1answer
143 views

How are new staining protocols designed for microscopy?

How are new staining protocols designed for microscopy? Is it manual work or does it rely on computer simulations? Say for example I thought the H&E staining does not bring enough contrast. How ...
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1answer
1k views

How long does it take to stain cells?

I'm never going to run these type of experiments but I do need to have a good idea of their timescale. After I fix my cells, if I stained my cells with histology stains like DAPI, Fluorescein, and ...
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1answer
664 views

What's a good reference for choosing histological staining chemicals?

It's often difficult to find the appropriate or best stain to use when I want to examine a new type of tissue. I think that's partly because many histological techniques were developed a long time ago,...
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2answers
2k views

Alternatives to ethidium bromide for staining small nucleic acids?

While ethidium bromide works well for staining larger single-stranded RNA or double-stranded DNA molecules, it doesn't stain smaller nucleic acids very well. I observed that at around 20 bases and ...
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535 views

Are these Gram stain substitutions acceptable?

In the context of a Gram stain on a blood smear: Are the following acceptable substitutions and/or what differences could arise by substituting them? Using methylene blue instead of crystal violet ...
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1answer
119 views

Is there a photobleaching-resistant, cell-permeant viability stain in the far red part of the spectrum?

I am looking for a live-cell–impermeable dye for viability. (The cells cannot be permeabilized and fixed in this experiment.) I would prefer with excitation and emission spectra similar to Cy5, but I ...
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1answer
760 views

To understand why satellite cells are genetically inactive in Barr body

DAPI is used as a stain for DNA heterochromatic and euchromatic regions. The Barr body is heterochromatic. In the slide of a normal human female cheek's somatic cells, there is apparently no other ...
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2answers
6k views

What is the Giemsa staining of chromosomes?

I got the question in my exam and wrote the following and I do not understand what is wrong in it: Giemsa staining is a staining method to stain particularly malaria and other parasital diseases. G-...
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116 views

Recommended applications for commercial antibodies

Commercial suppliers of primary antibodies for given protein targets typically list recommended applications, for which the antibody has presumably been shown to work. I am usually looking to use them ...
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3k views

What conditions should I use for Gel Red staining?

What are the optimal staining conditions when using Gel Red? So far, since we have started using it, the gels ran in our lab have been of very poor quality. The bands are very blurred and often ...
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1answer
289 views

What histological stain can I use for beta-keratin?

I'm trying to find a histochemical stain for beta-keratin, the type found in Reptilia which is organized in beta-sheets. It's different than alpha-keratin which is found in mammalian skin, hair, nails,...