The technique of using microscopes to view objects that are otherwise too small to be easily seen by the naked eye.

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choice of negative stains in general

I have isolated a large molecular complex whose integrity is very sensitive to increasing ionic strength and thus had to be prepared for EM using low ionic strength solutions. My goal is image this by ...
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Reasons preirradiation of negatively stained and plastic embedded specimens with a low electron dose improve stability to electron irradiation

It is known that preirradiation of negatively stained and plastic embedded specimens with a low electron dose improves their stability to electron irradiation. But my question is why this occurs? ...
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How to increase contrast with only a top entry specimen holder

My question is: If my microscope had a top entry specimen holder, instead of a side entry holder, what additional option(s) should I have to increase the contrast with the same constraint that I only ...
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16 views

a question about negative stains [duplicate]

I have isolated a large molecular complex whose integrity is very sensitive to increasing ionic strength and thus had to be prepared for EM using low ionic strength solutions. My goal is image this by ...
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19 views

background extraction in microscope analysis

I have taken microscope pictures for proteins in HeLa cells, in order to quantify the intensity (immunofluorescence). The proteins are both at the nuclear envelope, and in order to quantify the ...
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44 views

What microscope/magnification would I need to observe P. Acnes bacteria?

I am currently attempting to grow a culture of P. Acnes bacteria. Right now, my only hope in identifying colonies of the bacteria in the culture is to use a black light to find colonies that glow ...
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34 views

How can I reduce the background gradient in phase contrast microscopy?

I'm looking for ways to set up a phase contrast microscope to reduce the brightness gradient that appears across the background of my images. I use phase contrast to find the outlines of cells in my ...
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1answer
21 views

What Websites Have Image Libraries for Bacteria and other Microorganisms

What Websites Have Image Libraries for Bacteria and other Microorganisms? With age of cell phone microscopes and hand held spectrometers it would be interesting and valuable to be able to compare ...
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49 views

Is Zeiss Primo Start microscope worth the price for student use?

I study physics but I have got interested about biology too. Surfing around the web, I have found out that in buying microscopes one needs to decide between the cheaper no-name brands vs. the four big ...
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75 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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39 views

What are the features on a microscope one needs in order to do lab work?

By lab work I mean urinalysis, blood work(live as well), fecals, cytologies, histologies and all other. I have read(partly) a book(from 2002) on lab diagnostics and the author did not mention ...
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Slicing cell cultures frozen in vitrified ice with a laser for CryoEM

Could a cell culture frozen in vitrified ice for CryoEM be effectively sliced into thin planes and resolved images produced? I was thinking if this could be done to get images from inside a cell.
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79 views

Can one use a hemocytometer on a compound upright microscope?

I'm posting this as a follow-up on What to look for when buying a light microscope?. The answerer states that you would need to use a an inverted microscope to count cell in the hemocytometer(counting ...
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39 views

Can you find malaria parasites in saliva?

Can you find plasmodium parasites (malaria) in saliva under microscope from someone who's infected? Or it's only in the blood?
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62 views

How to visualize the ECM?

Specifically, I'd like to look at changes in HA (hyaluronic acid) production. Most often you only see people staining the cell surface or removing cells from culture for fixation and then imaging. ...
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1answer
17 views

Can inverted microscope also be phase contrast microscope?

I've read somewhere that inverted microscope also allows phase contrast microscopy. Is that true? Do all inverted microscope have this ability? Do they have any other functions? Many thanks in ...
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46 views

What is a good source that will help me classify protists?

I am a 15 year old that enjoys looking at the micro world. The only issue that I have is that I can't find a good online source that will help me classify the name of the micro organism that I'm ...
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40 views

Are there any drawbacks to having a fixed-Köhler microscope?

I am considering to buy the Zeiss Primo star microscope. There are two variants: one with so called "Full Köhler" illumination and one with "Fixed-Köhler" illumination. Are there any drawbacks to ...
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2answers
40 views

How much are phase-contrast and dark-field microscopy used in biological research? [closed]

How much are phase-contrast and dark-field microscopy used in biological research today? It seems these were invented a long time ago, so I am thinking would it be a good idea to learn these ...
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52 views

What are the dimensions in angstrom of bacteria? [closed]

I'm going to take a picture of a bacteria but I am unsure how large it is. What are the dimensions in angstrom of bacteria?
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306 views

Machine learning for light microscopy — problems to solve?

I would like solve some biological problems that would improve the state-of-the-art of biology or bioinformatics. In particular, I want to apply machine learning on light microscopic images. The ...
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1answer
83 views

Microscope scale bar

In some microscopic images, there is a bar scale at one of the corners or near a specific object. How are they added to the images? Does the slide have some markers, or are they digitally added? If ...
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1answer
50 views

Dataset of microscopic images before and after staining?

I am looking for a freely distributed dataset of microscopic images. I am hoping to find a dataset that would feature histological images before and after staining (e.g. H&E or some other standard ...
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1answer
297 views

Is it true that viruses cannot be seen with phase-contrast microscopy?

There appears to be a lot of material on the internet claiming that viruses can be only seen with electron microscopes, and not with light microscopes. To the contrary, for example this old paper ...
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58 views

How to operate a microtome?

In the picture above (Source: Amazon.com, United Scientific MCT001 Hand Microtome) we can see a microtome. How is it operated?
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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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263 views

What kind of microscope for ML/biological research?

I am a computer science student, focusing on machine learning applications. I have been always interested in biology but I lack any training in it. Now, I had an idea that I could introduce myself to ...
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122 views

Can oil be used instead of water in preparing wet mounts for microscopy?

Is there a reason that oil (e.g., vegetable or clear mineral) is not used to make wet mounts in microscopy? I am not referring to oil immersion technique but substituting oil for water in wet mounts ...
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33 views

Dyes to bind to double stranded DNA?

Are there any commercially available fluorescent dyes that will bind only to double stranded DNA (not RNA, single stranded DNA etc.) for studying in vitro using confocal microscopy?
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206 views

Benefits of CLARITY?

What are the benefits of CLARITY over this technique that was published more than a year earlier? Of course the second technique needs a fancier microscope that is likely more expensive and requires ...
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1answer
41 views

What's the best way to measure/calculate the size of a light beam at the sample of a microscope?

I am using a microscope with an LED derived light through the epi-fluorescent port of a microscope. I know that the "field of view" for a given objective is equal to the field number/magnification ...
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1answer
65 views

Mites on ginger root

I was just looking at some mouldy ginger found in the fridge under the microscope and was horrified/delighted to find (what looked to be) mites growing in it. I have searched the web and can only find ...
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1answer
77 views

What is this image?

What I understand is- I think this is picture of DNA. But why is it in this conformation ? What types of cells have this type of DNA ?
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53 views

Looking for raw cell videos from a light microscope

First off, I'm not looking for online lectures. I am trying to find creative commons licensed videos of live cells moving around without annotation or anything extra. I would prefer a few minutes of ...
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3answers
156 views

Assay for Beta-galactosidase activity in single cell microscopy

I'd like to be able to measure the activity of $\beta$-galactosidase in living cells with simple optical (maybe fluorescence) microscopy. Ideally I'd like to do a minimum of genetic engineering, and ...
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226 views

Image Processing Suite for bacterial microscopy: Schnitzcells or MicrobeTracker?

I am looking to start doing some work tracking the size and growth of individual bacterial cells in the microscope. In order to analyze the images I need software that can segment the cells, ...
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124 views

Image analysis of GFP-tagged protein localization bursts

I'm reading an article (full text here) that analyze the dynamics of localization of a GFP-tagged transcription factor (Crz1) over the time at the single-cell level, by taking movies in a fluorescent ...
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2answers
712 views

Microscope recommendation for an amateur?

I am a mathematics student looking for a hobby and this has come up as a potential hobby. Thus I am interested in buying a high-magnification microscope that is within the limits of my budget. My ...
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1answer
3k views

Isotropy of sarcomere bands in skeletal muscle cells

As seen under a polarization microscope, the A-Band in skeletal muscle fibres is so named because it is anisotropic in its refractive index which is a characteristic of orderly crystalline structure. ...
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61 views

Deducing synaptic strength from electron micrographs?

There are several promising techniques for connectomics based on iterative sectioning and imaging of tissue with scanning electron microscopes (e.g. FIB-SEM and ATUM) By looking at such micrographs, ...
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112 views

What are these stars in my microscopic image?

I see this very curious patterns on the sun-facing side of a jasmine (probably) leaf - visible with a makeshift microscope (about 50x) What are them? Pores? Are there other plants with similar ...
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1answer
129 views

Any difference between inverted microscope objective vs normal microscope objective?

I'm planning to build an inverted microscope and I am not sure if I can use a plane achromatic objective of a normal microscope for the inverted microscope as well. Are there any specific needs for ...
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1answer
254 views

Software for counting fly eggs

Is there any software that could be used to rapidly (quicker than humans) count the eggs laid by Drosophila on a substrate by placing them under a microscope with a camera attached? The eggs are laid ...
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1answer
223 views

What is a simple protocol for staining cells in suspension?

I am an engineering student studying how electric fields affect cells, specifically the phenomena of electroporation in living cells. I know that electroporation is widely used for introducing genes ...
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785 views

What kind of microscope should be used to view biological structures (such as spores) roughly 5 µm in length?

I would like to be able to identify genera of fungi based on the shape, size, and colour of spores. So the requirements are: Must be able to view a 5 micrometre object clearly with reasonable ...
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73 views

Microscopy Book Suggestions

I've learned programming through great book recommendations, many from the Stack Exchange series of sites. I'm hoping to take this approach to gaining a fundamental understanding of how fluorescence ...
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1answer
102 views

How to evaluate the efficacy of an antibody for fluorescence microscopy?

I'm trying to evaluate data taken from fluorescence microscopy with antibody staining, and am wondering whether there is any standard way to evaluate the specificity of the antibody for such ...
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91 views

What is the state of automated quantitative analysis?

Quantitative analysis of microscope images is essential for many types of studies, but the techniques used most often are very labor-intensive. I have been able to find some literature about using ...
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1answer
82 views

Atomic force microscopy: depth penetration?

What depth penetration can one typically get with atomic force microscopes? (i.e. how far into the tissue can one image?). I have found numbers on acquisition times (minutes) and spatial resolutions ...
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3answers
211 views

Super-resolution imaging in vivo

Are there any microscopy modalities or techniques for in vivo imaging at higher resolutions than the diffraction limit? I was looking at this list: Super resolution microscopy, but they don't talk ...