MICROSCOPE

Dr. Hajra Nazir

• LIGHT MICROSCOPE

• ELECTRON MICROSCOPE
Power/Magnification of lenses
• Eye piece lens = *10
• Low power OBJECTIVE lens = *10
• High power OBJECTIVE lens = *40
CARE WHEN USING HIGH POWER LENS
• High power lens is longer. So handle carefully, to avoid smashing the
lens or the slide.
• Focus by moving the lens away from the slide. Always focus upwards.
To avoid smashing.
 STAINED AND UNSTAINED SECTIONS
• No stain is used in unstained sections.
• Stains are used to provide a better colour contrast
• Commonly used stains are iodine (orange yellow) and methylene blue
PREPARING A SLIDE OF HUMAN CHEEK CELLS
• APPARATUS:
• light Microscope
• Slides and cover slips
• Cotton bud
• Forceps/mounting needle
• Dropping pipette
• Filter paper
• Methylene blue
• Beaker of disinfectant
• METHOD
• OBSERVATIONS:
- cytoplasm, nucleus, cell membrane seen
- Nucleus stains darker then cytoplasm
- Other Organelles not seen . ( as too small to be seen at this
magnification)
PRECAUTIONS
• Use a clean microscope slide
• Use a clean cotton bud
• Make sure scrapings are completely covered with the stain
• Avoid air bubble formation by gently lowering the cover slip at an
angle
• Place used cotton buds and slides in a disinfectant
• Wash hands after handling biological materials.
PREPARING A SLIDE OF ONION EPIDERMIS
• APPARATUS:
• Microscope
• Slides and cover slips
• Scalpel/razor
• Forceps
• Pipette
• Filter paper
• Iodine solution (orange yellow)
• Onion bulb (to examine onion epidermis)
METHOD:

• Cut a small piece out of onion leaf using a scalpel/razor

• Use forceps to peel a thin skin ( epidermis)of onion sample
• Place the epidermis flat on the slide using forceps
• Put one drop of water using pipette
• Place coverslip slowly at an angle of 45 degree, using forcep/mounting
needle, to avoid air bubbles.
• Put the slide on the stage of microscope and first view under low power
and then high power.
• Care to be taken when using high power lens.
• Observations??
• If preparing a stained section:
• Stain can be directly added to water when prepoaring the slide or can be
added later after viewing the unstained slide.
• Add a drop of stain (iodine) along one edge of cover slip using a pipette.
• Cover slowly with a cover slip at an angle toavoid air bubbles.
• Use filter paper/paper towel to help absorb excess stain .
• Put the slide on the stage of microscope and first view under low power
and then high power.
• Care to be taken when using high power lens.
• Observations:
- Cell wall, cell membrane and nucleus seen
- No chloroplasts visible
PRECAUTIONS :
• USE clean slide
• Take a thin section
• Place the peel FLAT on a wet slide, to avoid curling of the peel
• Place cover slip slowly and at an angle to avoid air bubbles
• Careful handling of iodine
UNSTAINED AND STAINED SECTION OF A GREEN PLANT CELL.

Iodine turns
purple/bluish
black in the
presence of starch.
Which are highly
concentrated in
the chloroplasts of
plant cells. Rest of
the cell stays
orang/brown.
Longitudnal section

Transverse section
• Hajra.nazir@alpha.edu.pk