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Lecture 2

lecture 2 - Imaging Cells .docx

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Biology 2382B
Robert Cumming

Lecture 2Imaging in Cell Biology Lecture 1looking at how we get the cells now we look at how we examine them imagining them Past Microscopes1600speople didnt realize existence of cellsAnton Van Leeuwenhoekin early 1600s was the first person to build a microscope that was good enough to see cells First to observe living protozoa and bacteria which he called animalculesWent onto visualize human red blood cells and spermWith great skill at grinding lensesnaturally acute eyesight and lots of patient he was able to achieve a magnification of 200X Modern Compound MicroscopeIn contrast to antons microscop this uses multiple lenses as opposed to one Brightfield microscopy o Light source which passes through a condenser to condense the light whichfocuses the light on the specimen o Objective lens then collects the light after it has passed through the specimen o Ocular or eyepiece lens to focus image onto the eyeo Typical light microscope magnification is 40 to 1000xo Only structures with a high refractice index ability to bend light are observable Light refractionthe ability to bend light or to slow it down Resolution of microscopesResolution the ability to distinguish between two very closely position objects as separate entities A conventional microscope can never resolve objectscellular features that are less than 02M Apart Smaller resolution is betterResolutionD Distance resolved between t points D061NsinWavelength of lightNsin Numerical aperture Higher is betterN Refractive index of medium between the specimen and the objective Lens angle of light entering objective lensThe limit of resolution is 02 m200 nm To make D smallerusing oil to increase the refractive index affect the angle based on the properties of how the lens is grounded or madeo Add some oil or water between the specimen to increase the refractive index
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