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Micro SDL PowerPoint-1

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6 views

Micro SDL PowerPoint-1

Uploaded by

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

Field
Microsco
Presented by:
Ananya Rajappa B R

Fig: A colony of g r e e n planktonic


alg ae Volvox under the dark fi eld
py (23SM1017) Aniket Sharma
(23SM1019)
Ann Theress (23SM1020)
Anna Thomas
microscope. Image Credit: D. (23SM1021) Anoushka
Kucharski K.
Singh (23SM1022)
History
In 1830, J.J. Lister (the father of Joseph Lister)
invented the dark fi eld microscope, in which the
standard bright field (Abbe) condenser is
replaced with a single or double- refl ecting dark
fi eld condenser.
In 1906 in Vienna, Karl Landsteiner and Viktor
Mucha were the first to use dark fi eld 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 b e stained by
standard methods, or are so distorted by staining
that their characteristics then cannot b e identifi ed.

In dark fi eld 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 fi eld condenser.
• The dark fi eld 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 refl ected by the
specimen enters the objective lens whereas the
unrefl ected 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 e d g e detail examination


Advantages
▶ Simple and cost-eff ective setup

▶ Ideal for Unstained, Transparent


Specimens

▶ Higher resolution and contrast

▶ No sample preparation needed

▶ Suitable for observing live organisms

▶ Detailed view of external features

▶ Eff ective for water-borne organisms


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

▶ Since the sample has to b e 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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