LAB

ASSIGNMENT:01

Fundamentals of Genetics
 Introduction to microscopy
.
 light microscope:
A light microscope (LM) is an instrument that uses visible light and
magnifying lenses to examine small objects not visible to the naked eye, or
in finer detail than the naked eye allows.

Figure 1 light microscope

Parts of light microscope:

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Figure 2 parts of light microscope(animated picture)

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 Figure 3 components of light microscope

Objective lens:

In microscopy, the objective lenses are the optical elements closest to the
specimen. The objective lens gathers light from the specimen, which is
focused to produce the real image that is seen on the ocular lens.

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 Figure 4 objective lens

Ocular lense:

An eyepiece, or ocular lens, is a type of lens that is attached to a variety of

optical devices such as telescopes and microscopes. It is so named because it
is usually the lens that is closest to the eye when someone looks through the
device

Figure 5 ocular lense

Magnification:

Magnification is the process of enlarging the apparent size, not physical size, of something. This
enlargement is quantified by a calculated number also called "magnification"

Formula

Magnification (m) = h / h’

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Here, h is the height of the object

h’ is the height of the object.

Resolution:

The resolution of an optical microscope is defined as the shortest distance

between two points on a specimen that can still be distinguished by the
observer or camera system as separate entities.

Stains used to visualize nucleus and DNA:

Hematoxylin - a nuclear stain that, with a mordant, stains nuclei blue-violet or
brown.

Hoechst stains - two types of fluorescent stains, 33258 and 33342, these are

to stain DNA in living cells.

Gram positive and Gram negative:

Gram-positive bacteria have a thick mesh-like cell wall made of peptidoglycan (50–90% of
cell envelope), and as a result are stained purple by crystal violet, whereas gram-negative
bacteria have a thinner layer (10% of cell envelope), so do not retain the purple stain and are
counter-stained pink by safranin.

Figure 6 gram negative

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 Figure 7 gram positive

LAB 4:

ELECTRON MICROSCOPE

An electron microscope is a microscope that uses a beam of accelerated electrons as a source of

illumination.It is a special type of microscope having a high resolution of images, able to magnify objects
in nanometres, which are formed by controlled use of electrons in vacuum captured on a
phosphorescent screen.

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 Figure 8 electron microscope

Types of Electron microscope:

1.The transmission electron microscope (TEM)

2. The scanning electron microscope (SEM)

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1.The transmission electron microscope (TEM):

 The transmission electron microscope is used to view thin specimens through which electrons
can pass generating a projection image.
 The TEM is analogous in many ways to the conventional (compound) light microscope.
 TEM is used, among other things, to image the interior of cells (in thin sections), the structure of
protein molecules (contrasted by metal shadowing), the organization of molecules in viruses and
cytoskeletal filaments (prepared by the negative staining technique), and the arrangement of
protein molecules in cell membranes (by freeze-fracture).

Figure 9 TEM

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2.The scanning electron microscope (SEM):

 Conventional scanning electron microscopy depends on the emission of secondary electrons

from the surface of a specimen.
 Because of its great depth of focus, a scanning electron microscope is the EM analog of a stereo
light microscope.
 It provides detailed images of the surfaces of cells and whole organisms that are not possible by
TEM. It can also be used for particle counting and size determination, and for process control.
 It is termed a scanning electron microscope because the image is formed by scanning a focused
electron beam onto the surface of the specimen in a raster pattern.

Figure 10 SEM