Confocal Scanning Micros

Confocal Scanning Microscopy was
patented by Marvin Minsky in the year

1957. After a few years, it was then
developed into a Confocal Laser Scanning
Microscope. The said microscope (CLSM) is
a combination of high resolution optical
imaging with depth selectivity that allows
us to perform optical sectioning. The CLSM
uses a laser beam instead of a lamp. It
works by passing a laser beam through a
light source aperture which is then focused
by an objective lens into a small area on the
surface of your sample and an image is built
up pixel-by-pixel by collecting the emitted
photons from the fluorophores in the
sample.
Reference:
https://bitesizebio.com/19958/what-is-confocal-laser-scanning-microscopy/
1. Enumerate the 4 magnifications of the microscope

 Red (scanning objective=4x)
 Yellow (LPO/ Low Power Objective= 10x)
 Blue (HPO/ High Power Objective= 40x)
 White (OIO/ Oil Immersion Objective=100x)
2. What are the principles of a bright field microscopy?

 The word bright field is derived from the fact that the specimen is dark & contrasted by
the surrounding bright viewing field. Simple light microscopes are also called sometimes
as bright field microscopes.
 They are said to be the first type of microscopes learned in schools.
 Bright field microscopy is easy to manipulate since there are fewer adjustments needed
to in order to view the sample specimen.
 Some studies like microbiology & bacteriology depend on the bright field technique.
3. Types of Microscope & their principles
1. OPTICAL MICROSCOPE
 Binocular Microscope-it allows easy observation of 3D objects at low magnification.
 Brightfield Microscope- it uses transmitted light to observe targets at high
magnification.
 Polarizing Microscope-it uses different light transmission characteristics of
materials, such as crystalline structures, to produce an image.
 Phase Contrast Microscope-it visualizes minute surface irregularities by using light
interference. Commonly used to observe living cells without staining them.
 Differential Interference Contrast Microscope- similar to the phase contrast, is used
to observe minute surface irregularities but at a higher resolution.
 Fluorescence Microscope- It is a biological microscope that observes fluorescence
emitted by samples by using special light sources such as mercury lamps. When
combined with additional equipment, bright field microscopes can also perform
fluorescence imaging
 Total Internal Reflection Fluorescence Microscope- A fluorescence microscope that
uses an evanescent wave to only illuminate near the surface of a specimen. The
region that is viewed is generally very thin compared to conventional microscopes.
 Laser Microscope (Laser Scanning Confocal Microscope)- it uses laser beams for
clear observation of thick samples with different focal distances.
 Multiphoton Excitation Microscope-The use of multiple excitation lasers reduces
damage to cells and allows high-resolution observation of deep areas. This type of
microscope is used to observe nerve cells and blood flow in the brain.
 Structured Illumination Microscope-A high-resolution microscope with advanced
technology to overcome limited resolution found in optical microscopes that are
caused by the diffraction of light.
2. ELECTRON MICROSCOPE
 There are two types of electron microscopes namely Transmission electron
microscope (TEM) and Scanning electron microscope (SEM), the two said
microscope both uses electron beams instead of light beams.
3. SCANNING PROBE MICROSCOPE (SPM)
 Under the scanning probe microscope we have the Atomic force microscope (AFM)
and the Scanning near-field optical microscope (SNOM), these two microscopes
scans the surface of samples with a probe and this interaction is used to measure
fine surface shapes or properties.
Other types of microscope include X-ray microscope, ultrasonic microscope, and etc.
4. Microscope maintenance, care & troubleshooting
MICROSCOPE CARE & USE
Handling Of The Microscope
 Always use two hands to move the microscope. Place one hand around the arm, lift
the scope, and then put your other hand under the base of the scope for support.
 Always have clean hands when handling your microscope.
Storing The Microscope
 Return the lowest power objective in place

 Wrap the cord around the base
 Dust is an enemy to microscope lenses; always keep the microscope covered when
not in use.
Cleaning the Microscope
 Don’t let the microscope get too dirty – always use the dust cover when not in
use.
 To clean the eyepiece – use a high quality lens paper. First brush any visible dust
from the lens, and then wipe the lens. Do not use facial tissues, they are made
from ground up wood fibers and could damage the lenses.
 To clean the objective lenses – use a fresh piece of the lens paper each time so
that you don't transfer dust from one lens to another.
 Use lens paper on all glass parts of the microscope.
 Clean oil immersion lens with chemicals provided by your instructor.
Reference:
https://www.microscopemaster.com/brightfield-microscopy.html
https://www.keyence.com/ss/products/microscope/bz-x/study/principle/type.jsp

Confocal Scanning Micros

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Confocal Scanning Microscopy was

patented by Marvin Minsky in the year

1. Enumerate the 4 magnifications of the microscope

2. What are the principles of a bright field microscopy?

Storing The Microscope

 Return the lowest power objective in place

Cleaning the Microscope

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