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Micro SDL PowerPoint-1

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7 views

Micro SDL PowerPoint-1

Uploaded by

23aniketsharma
Copyright
© © All Rights Reserved
Available Formats
Download as PDF, TXT or read online on Scribd
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Dark Field

Microscopy
Presented by:
Ananya Rajappa B R (23SM1017)
Aniket Sharma (23SM1019)
Ann Theress (23SM1020)
Fig: A colony of green planktonic algae Anna Thomas (23SM1021)
Volvox under the dark field microscope. Anoushka Singh (23SM1022)
Image Credit: D. Kucharski K. Anusha Ananda (23SM1023)
History
In 1830, J.J. Lister (the father of Joseph Lister) invented the
dark field microscope, in which the standard bright field
(Abbe) condenser is replaced with a single or double-
reflecting dark field condenser.
In 1906 in Vienna, Karl Landsteiner and Viktor Mucha were
the first to use dark field microscope to visualise T pallidum
from syphilis lesions.
What is Dark Field Microscopy?
Dark Field Microscopy is used to examine live micro-
organisms that are either invisible in the ordinary light
microscope, cannot be stained by standard methods, or are
so distorted by staining that their characteristics then
cannot be identified.

In dark field microscopy, the object appears bright against a


dark background.
Principle
In Dark Field (or dark ground) Microscope, the object
appears bright against a dark background. This is made
possible by use of a special dark field condenser.
• The dark field condenser has a central opaque area that
blocks light from entering the objective lens directly and
has a peripheral annular hollow area which allows the
light to pass through and focus on the specimen
obliquely.
• Only the light which is reflected by the specimen enters
the objective lens whereas the unreflected light does not
enter the objective. As a result, the specimen is brightly
illuminated; but the background appears dark.
Applications
 Enhanced contrast for transparent structures

 Visualisation of thin bacteria

 Rapid detection of Treponema pallidum

 Observation of Bacterial and Protozoan motility

 Study of marine organisms and materials like crystals and


polymers

 Examination of cells and tissues

 Surface and edge detail examination


Advantages
 Simple and cost-effective setup

 Ideal for Unstained, Transparent Specimens

 Higher resolution and contrast

 No sample preparation needed

 Suitable for observing live organisms

 Detailed view of external features

 Effective for water-borne organisms


Disadvantages
 Images formed are prone to degradation, distortion and
inaccuracies

 Since the sample has to be adequately illuminated,


sometimes strong illumination may damage the specimen

 Not only the specimen but dust and other particles


scatter the light and are easily observed

 Low Level of light and limited colours


Advancements
 Integration with Fluorescence Techniques

 Digital imaging and Real-time analysis

 Label-free and non-invasive imaging

 Portability and Accessibility

 Enhanced contrast for organisms without staining,


preserving their natural state

 Super Resolution Techniques extending visibility to


nanoscale structures
Thank you

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