Phase contrast and differential interference contrast (DIC) do not require stain

Question

Phase contrast and differential interference contrast (DIC) do not require staining.

3.Which of the following microscopy techniques can be used in live cell imaging?<br />1. Phase contrast <br />2. Differential Interference Contrast <br />3. Scanning electron microscopy<br />4. Immunoelectron microscop

fluorescence microscopy (0.2 µm), structured illumination microscopy (100 nm), stimulated emission depletion (30 nm), transmission electron microscopy (0.1 nm)

1. When studying a new protein, it is useful to generate an antibody against it to tag it in experiments designed to study its function and localization. Which of the following is NOT a step needed to generate this new antibody? Mouse spleen ß cells are fused with myeloma cells. Mice are injected with the antibody that will tag the new protein. Hybridoma cells are selected for their ability to divide and grow in a selection medium. The spleen is isolated from a laboratory mouse.

Explanation / Answer

1) When studying a new protein, it is useful to generate an antibody against it to tag it in experiments designed to study its function and localization. A step NOT needed to generate this new antibody is (b) Mice are injected with the antibody that will tag the new protein.

Here, the mice must not be injected with antibody that will tag the new protein, rather, it should be injected with the protein antigen only, in response to which a new repertoire of antibody against this antigen will be generated in the mice body.

2) With respect to microscopy, the statements which is NOT true is Option: Two proteins cannot be visualized simultaneously using fluorescence microscopy.

As in fluorescence microscopy, more than one protein can be visualized by tagging them with different fluorescent labels like GFP, YFP etc.

3) The microscopy techniques that can be used in live cell imaging are Phase contrast and Differential Interference Contrast microscopy.

Living cells can not be visualized with any kind of electron microscopy technique, as electron microscopy needs the cell to be fixed in a proper way before visualizing.

4) From higher resolution to lower resolution, the order will be (b) transmission electron microscopy (0.1 nm), stimulated emission depletion (30 nm), structured illumination microscopy (100 nm), fluorescence microscopy (0.2 µm).

The resolution of a microscope is defined as the shortest distance between two points of the specimen that can still be distinguished by the microscope.

Here, Transmission electron microscopy can be used to visualize the two points in the sample which are even 0.1 nm apart, whereas stimulated emission depletion can visualize the image of the specimen in which two points are of minimum 30 nm apart.

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