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Florence microscopy has 3 main different way to make the cell fluorescence each with the disadvantages.
Label protein outside the cell and microinject it - Takes a long time for little results
You can use Immunofloresence which is faster by direct or indirect. Indirect you get better results through more floresence and takes less time because you can order the antibodies off the internet.
You can use GFP gene and insert it into the cell. What you can do is have a plasmid transfect it then the protein will glow - This has a downside if the protein doesnt fold.
If you are doing the immunofloresence you need to prepare the cell. You need to kill it to use the antibodies. You use formaldhyde or gluteraldehyde or methanol for this. This will but everything in place (cytoplasm wont move) then use a detergent to punch holes in the membrane. Then you use your anti-bodies to go into the cell and attach to your primary proteins. that you want to see.
To make these antibodies you can proceed either by monoclonal or polyclonal. inject the rabit or w/e animal with an antigen. You will get antibodies forming with the epitop to that antigen that antigen should be the protein you want. Polyclonal have more then 1 epitope. and you need the rabit to do this.. if it dies bye bye antibodies disadvantage
Monoclonal have Hybridomas which are immortal cell lines, they regognize only 1 epitope.
Myeloma cell with a lymophocytes.
To make sure you only have there hybridoma cell you can put them in HAT which will kill the myloma cells and you only get hybridomas.
What else can I help you with?
What has the author F W D Rost written?
F. W. D. Rost has written: 'Quantitative fluorescence microscopy' -- subject(s): Fluorescence microscopy, Technique 'Fluorescence microscopy' -- subject(s): Fluorescence microscopy 'Photography with a microscope' -- subject(s): Photomicrography
What has the author H M Holz written?
H. M. Holz has written: 'Worthwhile facts about fluorescence microscopy' -- subject(s): Fluorescence microscopy
How effective is the dapi nuclear stain in visualizing cellular nuclei in fluorescence microscopy?
The dapi nuclear stain is highly effective in visualizing cellular nuclei in fluorescence microscopy.
What is the recommended mounting medium for fluorescence microscopy?
The recommended mounting medium for fluorescence microscopy is an anti-fade mounting medium, such as Vectashield or ProLong Gold, which helps preserve the fluorescence signal and prevent fading during imaging.
What is the definition of the term fluorescence microscopy?
Fluorescence microscopy is a technique used to visualize biological structures or molecules by inducing fluorescence in the sample and detecting the emitted light using specialized microscopes. This technique is commonly used in cell biology to study the localization and interaction of specific molecules within cells.
An advantage of fluorescence microscopy over staining techniques is that?
fluorescence microscopy allows for specific targeting of molecules or structures within a sample using fluorescent dyes or proteins, resulting in enhanced specificity and sensitivity compared to traditional staining techniques. Additionally, fluorescence microscopy enables dynamic imaging of live cells or tissues in real-time, providing insights into cellular processes and behaviors that cannot be captured by staining methods.
How does the use of a dichroic mirror enhance the performance of fluorescence microscopy?
A dichroic mirror enhances fluorescence microscopy by selectively reflecting and transmitting specific wavelengths of light. This allows for better separation of excitation and emission light, resulting in improved image quality and contrast in the final fluorescence image.
What methods are used for total internal reflections?
There are many methods. Like: Second harmonic imaging, 4Pi microscope, structured illumination and sarfus. Also, there are some fluorescence methods like: fluorescence microscopy and confocal microscopy.
Why is a mercury blub necessary for fluorescence microscopy?
A mercury bulb is necessary for fluorescence microscopy because it emits ultraviolet light, which is used to excite fluorescent molecules in the sample. When the fluorescent molecules absorb this light, they emit lower energy visible light, which is what is detected by the microscope to produce the fluorescence image.
What is an auramine?
An auramine is any of a family of fluorescent dyes used to stain tissues for fluorescence microscopy.
Why is DAPI used in fluorescence microscopy?
DAPI is used in fluorescence microscopy because it specifically binds to DNA, allowing researchers to visualize the location and distribution of DNA within cells. This helps in studying cell structure, chromosome organization, and cell division processes.
Beams of electrons that shine on fluorescent materials are used in?
fluorescence microscopy to excite fluorescence in the sample, allowing visualization of specific structures or molecules.
