ACTIVITY 1: THE

MICROSCOPE CONFOCAL
Team 2 - Olac . Opial . Perez

MICROSCOPE
OPERATING PRINCIPLE
Point illumination is the operating principle of a confocal
microscope. In simple terms, you can only see the
focused part of the specimen. The laser will scan the
focused part and the detector will measure the
illumination producing an image of the optical section.
The image is then collected in a computerized system as
data, forming a 3D image.

The main characteristic of the Confocal Microscope is

that it only detects what is focused and anything outside
the focus point, appears black.
ZEISS LSM 900, Mokobi, F. (2022). Confocal Microscope- Definition, Principle, Parts, Types, Labeled Diagram, Applications.
the confocal laser scanning microscope (CLSM) https://microbenotes.com/
 PARTS OF THE
CONFOCAL
MICROSCOPE
1. Objective lens
2. Out-of-focus plane
3. In-focus plane
4. Beam splitters
5. Detector
6. Confocal pinhole
(aperture)
7. Laser
8. Oscillator Mirrors
Mokobi, F. (2022). Confocal Microscope- Definition, Principle, Parts, Types, Labeled Diagram, Applications. Microscope | Types, Parts, History, Diagram, & Facts. (2023, January 5). Encyclopedia
https://microbenotes.com/ Britannica. https://www.britannica.com/technology/microscope/Confocal-microscopes
 IMAGE OF TISSUE CAPTURED
BY THE MICROSCOPE

Figure 1. 3D tissue volumes Figure 2. Confocal microscopy of muscular tissue in Figure 3. Confocal Microscopy of Fibroblast
Gallus gallus domesticus 96hpi with Yellow Fever cells
Real fluorescence microscopic view of human skin cells in culture.
17DD virus Nucleus are in blue, actin filaments are in red, tubulin was labeled
with green

Ermine, C. M., Chavez, C. A., & Thompson, L. H. (2021). Histological Manso, et al., (2016). Kinetic Study of Yellow Fever 17DD Viral Infection in Vshivkova.(2016). Confocal microscopy of fibroblast cells stock photo.
characterization and quantification of newborn cells in the adult rodent Gallus gallus domesticus Embryos. PLOS ONE, 11(5), e0155041. https://www.istockphoto.com/photo/confocal-microscopy-of-fibroblast-cells-
brain. STAR Protocols, 2(3), 100614. https://doi.org/10.1016/j.xpro.2021.100614 https://doi.org/10.1371/journal.pone.0155041 gm586706662-100704103
ADVANTAGES DISADVANTAGES
High spatial resolution Complexity and cost: expensive to
produce and acquire
Increased depth of field

Reduced photobleaching Limited penetration depth: not able

to image very deep within tissues
Reduced background noise
Low light sensitivity: less sensitive to
Live cell imaging low levels of light than other types of
Multiplexing capabilities microscopes

Accuracy: no interference from scattered Limited field of view: have a small

light from other portions of the specimen field of view compared to other
types of microscopes
Living and Dead Cell: can be used to
examine both living and dead cells
Produces 3D picture sets
MN Editors. (2022, December 31). Confocal Microscope Principle, Uses, Parts, Advantages, And Disadvantages. Microbiology Note –
Online Biology Notes. https://microbiologynote.com/confocal-microscope-principle-uses-parts-advantages-and-disadvantages/