power point discusses basic information about the advent and use of light
microscopy. The history of the advent of microscopy dated back to 14th century,
several scientist contributed to the discovery and application of microscope,
the art of grinding lenses was developed in Italy and spectacles were made to
improve eyesight by Z. Janesen. One important point about the application of light
microscope in quantitative analysis of specimen is detail understanding of
sources of light and how they interact with the specimen of choice.
are some properties (such as type of sample, dimensions, the resolution,
demarcation, image area etc.) that determines the outcome of quantitative
microscopy which depends on the objective of the researcher. It is also
important to reduce some effects such as photobleaching and phototoxicity.
There are two different ways of imaging samples which are transmitted light
microscopy and reflected light microscopy. Reflected light microscopy is
applied when imaging specimen which do not transmit light as opposed to
transmitted light microscopy. There are several methods for the preparation of
sample which includes: immunohistochemical staining for non-fluorescent
contrast enhancement, Fluorescence–excitation with one (or more) wavelength and
light emitted with different wavelength is collected, Fixation–fixed cell
samples, Immunofluorescence with antibody staining. There are two important
properties in microscopy that defines the function of optics important in
microscopy, the objective lens which controls magnification and numerical
aperture (the light gathering ability) determines brightness and resolving power.
Another properties important in microscopy is resolution, which is the least
detectable detail in the image and a major factor that determines the quality
of image seen under the microscope. There are some factors that can affect the
resolution of a microscope. They are the following; numerical aperture and
aberration of objective, immersion, mounting medium, and cover glass used for
specimen, excitation wavelength, and colour range which affects the lighting
and fineness details, and system stability.
has advanced from quantitative where image is analyzed based on quantify,
counting large image set to automated
image understanding where computerized mage interpretation by automatizing
image analysis helps in quantifying large image sets.