The document provides an overview of fluorescence microscopy techniques, highlighting its various applications such as determining protein localization, studying dynamics, and examining cellular structures. It discusses different types of light sources and the basic design of fluorescence microscopes. The document also mentions various filter and objective lens configurations used in microscopy.

1. Microscope-III
Md. Saiful Islam
B.Pharm, M.Pharm (PCP)
North South University
Fb Group: Pharmacy Universe

2. A reminder…
• Brightfield - absorption
• Darkfield - scattering
• Phase Contrast - phase interference
• Fluorescence Contrast - fluorescence

3. What can you do with a fluorescence microscope?
•Determine the localisation of specific (multiple) proteins
•Determine the shape of organs, cells, intracellular
structures
•Examine the dynamics of proteins
•Study protein interactions or protein conformation
•Examine the ion concentration.
can observe in live cells

4. upright microscope light path

5. upright microscope light path
Lamp
Filter cube
Objective lens
sample
Camera
For bright field microscopy

6. Basic design of fluorescence
Objective acts as condenser; excitation light reflected away from eyes

7. Light source for fluorescence microscopy
Arc lamps
Xenon
Mercury
UV IR
Laser types
Argon 351 364 457 477 488 514
Blue diode 405 440
Helium-Cadmium 354 442
Krypton-Argon 488 569 647
Green Helium-Neon 543
Yellow Helium-Neon 594
Orange Helium-Neon 612
Red Helium-Neon 633
Red diode 635 650
Ti:Sapphire 720-980