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Fluorescence Microscopy vs Flow Cytometry


Measuring Cells vs Measuring Cells in Suspension
microscope vs flow cytometer
                           Fluorescence Microscopy and Flow Cytometry, both are biological instruments which are capable of measuring the biophysical and biochemical properties of cells or particles which are under study. Both of them are capable of identifying types of cells present in a given sample. These two technologies are useful in variety of applications and they have widespread usage.. But, These two differ in their applications. 

Fluorescence Microscopy:  Microscope was invented before flow cytometry, fluorescence microscope is improved version of a light microscope, uses the lasers as excitation sources to excite the fluorescence present in the specimen under study. Fluorescence emitted from the cells or particles present in specimen produces a magnified image. Fluorescence microscope is useful to study the specific the cell characteristics. With the help of this, cells can be quantified and can be analyzed qualitatively. However, number of cells or types of cells are limited. 

Flow Cytometry: with the help of this technology, cells or particles present in a suspension cab be quantified and analyzed & perform measurements for the characteristics such as cell size, granularity, and quantity of DNA or RNA, fluorescence and intracellular proteins..  Results obtained from the flow cytometer could be absolute or relative data. This has widely used for the identification and characterization of cells, cell types, counting, and identifying various extra cellular and intra cellular protein markers, identification and characterization of microorganisms. 



Fluorescence Microscope VS Flow Cytometer: 

Fluorescence Microscope and Flow Cytometer, both are capable of measuring the cellular components quantitatively. But, they differ in how many cells they can handle, locating the cellular components, volume of the cells, number of parameters and also sample preparation. 

Fluorescence Microscope can show how the cellular components are distributed inside the cell (clustered or uniformly). It can also show the area of the cell and its structures. It can quantify the cellular components for 10-100s of cell and one parameter at a time. This technology produces magnified images of the cells. Sample preparation involves the mounting the specific number of cells on a slide and observe under the microscope to produce a magnified image. One fluorescence parameter can be used simultaneously. Can not sort the cells. 

But, 

Flow cytometer can not show the distribution of the cellular components within the cell. But, it can measure the fluorescence tagged with those cellular components and measure them quantitatively. It can give relative data of the cell size, cell granularity and other properties such as fluorescence. It can handle up to 1000s of cells per second. (There is no magnified image.) Sample preparation involves the preparation of single cell suspension and maintaining these cells dispersed in the suspension. Up to 15-24 fluorescence parameters can be used simultaneously. Also, target cells under study can be sorted using the technique of fluorescence activated cell sorting and use these sorted cells for further experiments such as western blotting, RNA or DNA isolation or cell culturing. 
"Flow Cytometer and Fluorescence Microscope achieve same goals. They have their own roles, benefits, advantages and applications. It makes them suitable for the different specific roles".


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