SlideShare a Scribd company logo
by
c.Keerthana
 First

described by Dutch physicist frits
Zernike in 1934.
 It is a type of light microscopy.
 It is a contrast enhancing optical technique
that produces high contrast images of
transparent specimens.
 Specimen- unstained and alive.
 ANNULAR

RING : is between condenser and
light source.
 PHASE RING : is between objective lens and
image plane.
 SPL PROPERTY : both the rings allow partial
light to pass through it and the rest is
blocked.
Basic mechanism is interference of light
beams.
 INTERFENRENCE:
Interaction of two light waves which leads to
the formation of resultant wave.
 TYPES OF INTERFERENCE:
constructive interference
destructive interference
Light source
Annular ring
Condenser
Specimen plate
(interference)
objective lens

phase ring
Image plane
 Light

passes through the condenser via
annular ring.
 After reaching the specimen plate two types
of beams are formed.
 IF THERE IS NO SPECIMEN IN LENS:
1.Surrounding wave (S)
2.particle wave (p)
P=S
NO INTERFERENCE
 IF

LENS CONTAINS SAMPLE:
 Light beam gets diffracted because of
different density at different regions of
sample.
1.surrounding wave (S)
2.diffaracted wave (D)
P=S+D
Either constructive interference or
destructive interference may occur.
 Positive

phase contrast produces
Constructive interference.
Thus, the image of the specimen obtained is
 Inner region of the sample – darker
 Outer region of the sample– bright
 Surrounding lens – opaque
 Negative

phase contrast microscopy produces
destructive interference.
 Thus, the image obtained is
 Inner region of the sample – bright
 Outer region of the sample– darker
 Surrounding lens – opaque
 Fluorescence

microscope is
an one of the light microscope.
 It refers to any microscope
that uses fluorescence to
generate an image.
 It produces 3d image.
 The technique is used to
study specimens, which
can be made to fluorescence.
 Fluorescence

is a
phenomenon that takes place
when a substance absorbs
light at a given wavelength
and emits light at another
wavelength.
 Fluorescence occurs as an
electron, which has been
excited to a higher, and more
unstable energy state,
relaxes to its ground state
and gives off a photon of
light.
 The

sample to be analyzed Is placed on a
lens. And the sample is coated with a
fluorescence material.
 The light is illuminated through the lens with
the higher energy source. The illumination
light is absorbed by the fluorophores.
 The sample causes them to emit a longer
lower energy wavelength light.
 This fluorescent light can be separated from
the surrounding radiation with filters.
The light from the light
source is passed through
the excitation filter.
 The specific wavelength of
light is passed through the
sample via dichronic filter.
 The objective lens focuses
the light to the specimen.
 The light emitted from the
specimen is filtered by
barrier filter.

 Imaging

structural components of small
specimens, such as cells.
 Conducting viability studies on cell
populations (are they alive or dead).
 Imaging the genetic material within a cell
(DNA and RNA).
 Viewing specific cells within a larger
population with techniques such as FISH.
 To differentiate different type of cell.
Light phase contrast and fluorescence microscopy

More Related Content

What's hot

What's hot (20)

phase contrast microscope
phase contrast microscopephase contrast microscope
phase contrast microscope
 
Phase contrast microscope
Phase contrast microscopePhase contrast microscope
Phase contrast microscope
 
DARK FIELD MICROSCOPY
DARK FIELD MICROSCOPYDARK FIELD MICROSCOPY
DARK FIELD MICROSCOPY
 
Phase contrast & Flurosence microscope
Phase contrast & Flurosence microscopePhase contrast & Flurosence microscope
Phase contrast & Flurosence microscope
 
Principles and application of light, phase constrast and fluorescence microscope
Principles and application of light, phase constrast and fluorescence microscopePrinciples and application of light, phase constrast and fluorescence microscope
Principles and application of light, phase constrast and fluorescence microscope
 
Bright field microscope and its working principle
Bright field microscope and its working principleBright field microscope and its working principle
Bright field microscope and its working principle
 
Confocal microscopy by prabhat
Confocal microscopy by prabhatConfocal microscopy by prabhat
Confocal microscopy by prabhat
 
Confocal microscope
Confocal microscopeConfocal microscope
Confocal microscope
 
Fluorescence microscopy
Fluorescence microscopyFluorescence microscopy
Fluorescence microscopy
 
Fluoresence microscope
Fluoresence microscopeFluoresence microscope
Fluoresence microscope
 
FLUORESCENCE MICROSCOPY
FLUORESCENCE MICROSCOPYFLUORESCENCE MICROSCOPY
FLUORESCENCE MICROSCOPY
 
Fluorescence Microscopy
Fluorescence MicroscopyFluorescence Microscopy
Fluorescence Microscopy
 
Fluorescence microscopy
Fluorescence microscopyFluorescence microscopy
Fluorescence microscopy
 
Fluorescence microscope
Fluorescence microscopeFluorescence microscope
Fluorescence microscope
 
Dark Field microscopy
Dark Field microscopyDark Field microscopy
Dark Field microscopy
 
Differential Interference Contrast Microscope.pptx
Differential Interference Contrast Microscope.pptxDifferential Interference Contrast Microscope.pptx
Differential Interference Contrast Microscope.pptx
 
Phase contrast microscope
Phase contrast microscopePhase contrast microscope
Phase contrast microscope
 
Phase contrast microscope
Phase contrast microscopePhase contrast microscope
Phase contrast microscope
 
DARK FIELD MICROSCOPY
DARK FIELD MICROSCOPYDARK FIELD MICROSCOPY
DARK FIELD MICROSCOPY
 
Phase contrast microscope
Phase contrast microscopePhase contrast microscope
Phase contrast microscope
 

Viewers also liked

Fluorescence spectroscopy
Fluorescence spectroscopyFluorescence spectroscopy
Fluorescence spectroscopy
Nimisha Dutta
 
Market Research Report : Lighting industry in india 2015 - Sample
Market Research Report : Lighting industry in india 2015 - SampleMarket Research Report : Lighting industry in india 2015 - Sample
Market Research Report : Lighting industry in india 2015 - Sample
Netscribes, Inc.
 

Viewers also liked (15)

Phase contrast microscopy
Phase contrast microscopyPhase contrast microscopy
Phase contrast microscopy
 
Fluorescence Microscopy
Fluorescence MicroscopyFluorescence Microscopy
Fluorescence Microscopy
 
Fluorescence microscopy introduction
Fluorescence microscopy introductionFluorescence microscopy introduction
Fluorescence microscopy introduction
 
Microscopy
MicroscopyMicroscopy
Microscopy
 
Fluorescence spectroscopy
Fluorescence spectroscopyFluorescence spectroscopy
Fluorescence spectroscopy
 
Miniaturized light sheet based fluorescence microscopy Schneckenburger
Miniaturized light sheet based fluorescence microscopy SchneckenburgerMiniaturized light sheet based fluorescence microscopy Schneckenburger
Miniaturized light sheet based fluorescence microscopy Schneckenburger
 
White Paper: In vivo Fiberoptic Fluorescence Microscopy in freely behaving mice
White Paper: In vivo Fiberoptic Fluorescence Microscopy in freely behaving miceWhite Paper: In vivo Fiberoptic Fluorescence Microscopy in freely behaving mice
White Paper: In vivo Fiberoptic Fluorescence Microscopy in freely behaving mice
 
LED v Fluorescent - all you need to know
LED v Fluorescent - all you need to knowLED v Fluorescent - all you need to know
LED v Fluorescent - all you need to know
 
Rad 104 hospital practice and care of patients 3 drugs & contrast 2016
Rad 104 hospital practice and care of patients 3  drugs & contrast 2016Rad 104 hospital practice and care of patients 3  drugs & contrast 2016
Rad 104 hospital practice and care of patients 3 drugs & contrast 2016
 
How to Segment a Market
How to Segment a MarketHow to Segment a Market
How to Segment a Market
 
Lecture 11 systems of nonlinear equations
Lecture 11 systems of nonlinear equationsLecture 11 systems of nonlinear equations
Lecture 11 systems of nonlinear equations
 
Application of Fluorescence Activated-Cell Sorting (FACS) in separation of di...
Application of Fluorescence Activated-Cell Sorting (FACS) in separation of di...Application of Fluorescence Activated-Cell Sorting (FACS) in separation of di...
Application of Fluorescence Activated-Cell Sorting (FACS) in separation of di...
 
Bright field microscopes
Bright field microscopesBright field microscopes
Bright field microscopes
 
04 the microscope
04 the microscope04 the microscope
04 the microscope
 
Market Research Report : Lighting industry in india 2015 - Sample
Market Research Report : Lighting industry in india 2015 - SampleMarket Research Report : Lighting industry in india 2015 - Sample
Market Research Report : Lighting industry in india 2015 - Sample
 

Similar to Light phase contrast and fluorescence microscopy

microscopy in bacteriology by Tejpal.pptx
microscopy in bacteriology by Tejpal.pptxmicroscopy in bacteriology by Tejpal.pptx
microscopy in bacteriology by Tejpal.pptx
Vetico
 
2 Microscopy and staining.pptx
2 Microscopy and staining.pptx2 Microscopy and staining.pptx
2 Microscopy and staining.pptx
NeelGamers
 
Unit – 2 Microscopy
Unit – 2 MicroscopyUnit – 2 Microscopy
Unit – 2 Microscopy
sanchu yadav
 
Optical properties of nanomaterials
Optical properties of nanomaterialsOptical properties of nanomaterials
Optical properties of nanomaterials
udhay roopavath
 
Fluorimetry.pptx
Fluorimetry.pptxFluorimetry.pptx
Fluorimetry.pptx
Ankita Raikwar
 

Similar to Light phase contrast and fluorescence microscopy (20)

Microscopy
MicroscopyMicroscopy
Microscopy
 
microscopy in bacteriology by Tejpal.pptx
microscopy in bacteriology by Tejpal.pptxmicroscopy in bacteriology by Tejpal.pptx
microscopy in bacteriology by Tejpal.pptx
 
FLUOROSCENCE & PHASE CONTRAST MICROSCOPY.pptx
FLUOROSCENCE & PHASE CONTRAST MICROSCOPY.pptxFLUOROSCENCE & PHASE CONTRAST MICROSCOPY.pptx
FLUOROSCENCE & PHASE CONTRAST MICROSCOPY.pptx
 
Phase contrast and Fluorescent microscope
Phase contrast and Fluorescent microscopePhase contrast and Fluorescent microscope
Phase contrast and Fluorescent microscope
 
Microscope ppt, by jitendra kumar pandey,medical micro,2nd yr, mgm medical co...
Microscope ppt, by jitendra kumar pandey,medical micro,2nd yr, mgm medical co...Microscope ppt, by jitendra kumar pandey,medical micro,2nd yr, mgm medical co...
Microscope ppt, by jitendra kumar pandey,medical micro,2nd yr, mgm medical co...
 
MICROSCOPY.pptx
MICROSCOPY.pptxMICROSCOPY.pptx
MICROSCOPY.pptx
 
Types and applications of Microscopy.ppt
Types and applications of Microscopy.pptTypes and applications of Microscopy.ppt
Types and applications of Microscopy.ppt
 
Fluorimetry ANALYSIS- SPECTOSCOPY PPT
Fluorimetry ANALYSIS- SPECTOSCOPY PPTFluorimetry ANALYSIS- SPECTOSCOPY PPT
Fluorimetry ANALYSIS- SPECTOSCOPY PPT
 
Microscopy and Micrometry
Microscopy and MicrometryMicroscopy and Micrometry
Microscopy and Micrometry
 
Ppt f1
Ppt f1Ppt f1
Ppt f1
 
2 Microscopy and staining.pptx
2 Microscopy and staining.pptx2 Microscopy and staining.pptx
2 Microscopy and staining.pptx
 
Unit – 2 Microscopy
Unit – 2 MicroscopyUnit – 2 Microscopy
Unit – 2 Microscopy
 
microscopy
 microscopy microscopy
microscopy
 
Scanning and Transmission Electron Microscopy
Scanning and Transmission Electron MicroscopyScanning and Transmission Electron Microscopy
Scanning and Transmission Electron Microscopy
 
Optical properties of nanomaterials
Optical properties of nanomaterialsOptical properties of nanomaterials
Optical properties of nanomaterials
 
Fluorimetry.pptx
Fluorimetry.pptxFluorimetry.pptx
Fluorimetry.pptx
 
Microscopy presentation
Microscopy presentationMicroscopy presentation
Microscopy presentation
 
Florescense microscopy
Florescense microscopyFlorescense microscopy
Florescense microscopy
 
Fluorescence microscopy
Fluorescence microscopy   Fluorescence microscopy
Fluorescence microscopy
 
Light
LightLight
Light
 

Recently uploaded

IATP How-to Foreign Travel May 2024.pdff
IATP How-to Foreign Travel May 2024.pdffIATP How-to Foreign Travel May 2024.pdff
IATP How-to Foreign Travel May 2024.pdff
17thcssbs2
 
The basics of sentences session 4pptx.pptx
The basics of sentences session 4pptx.pptxThe basics of sentences session 4pptx.pptx
The basics of sentences session 4pptx.pptx
heathfieldcps1
 
Industrial Training Report- AKTU Industrial Training Report
Industrial Training Report- AKTU Industrial Training ReportIndustrial Training Report- AKTU Industrial Training Report
Industrial Training Report- AKTU Industrial Training Report
Avinash Rai
 

Recently uploaded (20)

Morse OER Some Benefits and Challenges.pptx
Morse OER Some Benefits and Challenges.pptxMorse OER Some Benefits and Challenges.pptx
Morse OER Some Benefits and Challenges.pptx
 
Instructions for Submissions thorugh G- Classroom.pptx
Instructions for Submissions thorugh G- Classroom.pptxInstructions for Submissions thorugh G- Classroom.pptx
Instructions for Submissions thorugh G- Classroom.pptx
 
IATP How-to Foreign Travel May 2024.pdff
IATP How-to Foreign Travel May 2024.pdffIATP How-to Foreign Travel May 2024.pdff
IATP How-to Foreign Travel May 2024.pdff
 
PART A. Introduction to Costumer Service
PART A. Introduction to Costumer ServicePART A. Introduction to Costumer Service
PART A. Introduction to Costumer Service
 
Jose-Rizal-and-Philippine-Nationalism-National-Symbol-2.pptx
Jose-Rizal-and-Philippine-Nationalism-National-Symbol-2.pptxJose-Rizal-and-Philippine-Nationalism-National-Symbol-2.pptx
Jose-Rizal-and-Philippine-Nationalism-National-Symbol-2.pptx
 
Students, digital devices and success - Andreas Schleicher - 27 May 2024..pptx
Students, digital devices and success - Andreas Schleicher - 27 May 2024..pptxStudents, digital devices and success - Andreas Schleicher - 27 May 2024..pptx
Students, digital devices and success - Andreas Schleicher - 27 May 2024..pptx
 
The Art Pastor's Guide to Sabbath | Steve Thomason
The Art Pastor's Guide to Sabbath | Steve ThomasonThe Art Pastor's Guide to Sabbath | Steve Thomason
The Art Pastor's Guide to Sabbath | Steve Thomason
 
50 ĐỀ LUYỆN THI IOE LỚP 9 - NĂM HỌC 2022-2023 (CÓ LINK HÌNH, FILE AUDIO VÀ ĐÁ...
50 ĐỀ LUYỆN THI IOE LỚP 9 - NĂM HỌC 2022-2023 (CÓ LINK HÌNH, FILE AUDIO VÀ ĐÁ...50 ĐỀ LUYỆN THI IOE LỚP 9 - NĂM HỌC 2022-2023 (CÓ LINK HÌNH, FILE AUDIO VÀ ĐÁ...
50 ĐỀ LUYỆN THI IOE LỚP 9 - NĂM HỌC 2022-2023 (CÓ LINK HÌNH, FILE AUDIO VÀ ĐÁ...
 
The basics of sentences session 4pptx.pptx
The basics of sentences session 4pptx.pptxThe basics of sentences session 4pptx.pptx
The basics of sentences session 4pptx.pptx
 
Benefits and Challenges of Using Open Educational Resources
Benefits and Challenges of Using Open Educational ResourcesBenefits and Challenges of Using Open Educational Resources
Benefits and Challenges of Using Open Educational Resources
 
aaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa
aaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa
aaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa
 
How to Create Map Views in the Odoo 17 ERP
How to Create Map Views in the Odoo 17 ERPHow to Create Map Views in the Odoo 17 ERP
How to Create Map Views in the Odoo 17 ERP
 
NCERT Solutions Power Sharing Class 10 Notes pdf
NCERT Solutions Power Sharing Class 10 Notes pdfNCERT Solutions Power Sharing Class 10 Notes pdf
NCERT Solutions Power Sharing Class 10 Notes pdf
 
How to Manage Notification Preferences in the Odoo 17
How to Manage Notification Preferences in the Odoo 17How to Manage Notification Preferences in the Odoo 17
How to Manage Notification Preferences in the Odoo 17
 
An Overview of the Odoo 17 Discuss App.pptx
An Overview of the Odoo 17 Discuss App.pptxAn Overview of the Odoo 17 Discuss App.pptx
An Overview of the Odoo 17 Discuss App.pptx
 
Sha'Carri Richardson Presentation 202345
Sha'Carri Richardson Presentation 202345Sha'Carri Richardson Presentation 202345
Sha'Carri Richardson Presentation 202345
 
The Benefits and Challenges of Open Educational Resources
The Benefits and Challenges of Open Educational ResourcesThe Benefits and Challenges of Open Educational Resources
The Benefits and Challenges of Open Educational Resources
 
Danh sách HSG Bộ môn cấp trường - Cấp THPT.pdf
Danh sách HSG Bộ môn cấp trường - Cấp THPT.pdfDanh sách HSG Bộ môn cấp trường - Cấp THPT.pdf
Danh sách HSG Bộ môn cấp trường - Cấp THPT.pdf
 
Open Educational Resources Primer PowerPoint
Open Educational Resources Primer PowerPointOpen Educational Resources Primer PowerPoint
Open Educational Resources Primer PowerPoint
 
Industrial Training Report- AKTU Industrial Training Report
Industrial Training Report- AKTU Industrial Training ReportIndustrial Training Report- AKTU Industrial Training Report
Industrial Training Report- AKTU Industrial Training Report
 

Light phase contrast and fluorescence microscopy

  • 2.  First described by Dutch physicist frits Zernike in 1934.  It is a type of light microscopy.  It is a contrast enhancing optical technique that produces high contrast images of transparent specimens.  Specimen- unstained and alive.
  • 3.
  • 4.  ANNULAR RING : is between condenser and light source.  PHASE RING : is between objective lens and image plane.  SPL PROPERTY : both the rings allow partial light to pass through it and the rest is blocked.
  • 5. Basic mechanism is interference of light beams.  INTERFENRENCE: Interaction of two light waves which leads to the formation of resultant wave.  TYPES OF INTERFERENCE: constructive interference destructive interference
  • 6.
  • 7. Light source Annular ring Condenser Specimen plate (interference) objective lens phase ring Image plane
  • 8.  Light passes through the condenser via annular ring.  After reaching the specimen plate two types of beams are formed.  IF THERE IS NO SPECIMEN IN LENS: 1.Surrounding wave (S) 2.particle wave (p) P=S NO INTERFERENCE
  • 9.  IF LENS CONTAINS SAMPLE:  Light beam gets diffracted because of different density at different regions of sample. 1.surrounding wave (S) 2.diffaracted wave (D) P=S+D Either constructive interference or destructive interference may occur.
  • 10.  Positive phase contrast produces Constructive interference. Thus, the image of the specimen obtained is  Inner region of the sample – darker  Outer region of the sample– bright  Surrounding lens – opaque
  • 11.  Negative phase contrast microscopy produces destructive interference.  Thus, the image obtained is  Inner region of the sample – bright  Outer region of the sample– darker  Surrounding lens – opaque
  • 12.  Fluorescence microscope is an one of the light microscope.  It refers to any microscope that uses fluorescence to generate an image.  It produces 3d image.  The technique is used to study specimens, which can be made to fluorescence.
  • 13.
  • 14.  Fluorescence is a phenomenon that takes place when a substance absorbs light at a given wavelength and emits light at another wavelength.  Fluorescence occurs as an electron, which has been excited to a higher, and more unstable energy state, relaxes to its ground state and gives off a photon of light.
  • 15.  The sample to be analyzed Is placed on a lens. And the sample is coated with a fluorescence material.  The light is illuminated through the lens with the higher energy source. The illumination light is absorbed by the fluorophores.  The sample causes them to emit a longer lower energy wavelength light.  This fluorescent light can be separated from the surrounding radiation with filters.
  • 16. The light from the light source is passed through the excitation filter.  The specific wavelength of light is passed through the sample via dichronic filter.  The objective lens focuses the light to the specimen.  The light emitted from the specimen is filtered by barrier filter. 
  • 17.  Imaging structural components of small specimens, such as cells.  Conducting viability studies on cell populations (are they alive or dead).  Imaging the genetic material within a cell (DNA and RNA).  Viewing specific cells within a larger population with techniques such as FISH.  To differentiate different type of cell.