© Microscopy is a scientific technique that involves the use of Microscope 40 view the objects ‘thot are too small to be seen with noked eyes © A Microscope 1S an instrument ushich consist of lens or ombinalion of lenses used to magnify the objects that cannot be seen with Naked eyes oT wlitses light Ceither sunlight or artificial light) 0 elechon beam to see the enlarged images of objectoy mt unnggany we see the enlamged images of object In_Microscopes. ‘ope mainly consist oF two types OF Lenses A Mi Oailar Lenses Eye Plee) Ihjedive Lenses, Components OF MicroscoPe Eve Pr - am Banu ThorComponents OF Microscope Eve Piece Coarse Aowsment Kyos Booy Tuse Fine Aowust¥eNT Nos Revowwing, Nose Piece Obsectve Lens Arm Convenser Twcunanion Jom Minor BaseMAGNIFICATION © Fist Magniftation > Objedve Lens © Send Magnification —* Ocular Lens © Final Magnification > lala od Sx) RESOWTON. © Th microscopy , fesolution refer, to the ability of a microscope to distinquigh fwo points aS separate entities © Higher resolution mena finer deals OF specimen can be cbseaved© mgner resolunon menng fier detauls Ot Specimen can ve” onserUed Tyees OF Microscopy Microscopy of mainly two typs © Light Microscopy @ Becton Microscopy MicroscoPe Light Microscope. Phaco (Anknel Micmnemne Scnina Electron© Light Microscopy @ Becton Microscopy Microscope Phase Gntrast Microscope Scanning, Electron Dak Field Microscope. Microscope Bright Field Microscope Transmission Eledron Fluoroscence Microscope Microscope.Puase Contrast Microscope @ This Micoscope was invented by. a Dutch Physicist Faits Zemike in 1934. He wos algo worded” by. Nobel Prize in 1953 for this © F ig a microscope that allowa observation of lining, cell eo Tt is 0 type of micosmpe that allowa the obsesvation of Frangpasents Objects ike ving, lls by. Converting, phate shifts In light @ enhancing, @ntrast @ With the hep of Nise Contrast microscope ,we Can (entity frangparent object's without Shaining, unlike Bright Fekel MicroscopyThe Prin Phase Conlrast Microscopy involues Conversion of smal phase changes in light rays, induced by dliferences in thickness index of different pasts of an Object , into visible anges in brightness Condenser Specimen Objective Phase Plate. ‘Annular Retarded _Unretarded| Diaphragm Rays RaysKey Components & WoORkING MeqianisM Light Source 2 The microscope uses a bright light sowre , typically @ Halogen or Led Lamp to Muminole the specimen Annular Diaphragm]: IF is locoted tn the ndenser and adjust the Phase Plate. light 40. ome foom all directions, . + JF is located in objective leng The phase plate inroduwes a phase shift to the light thot passes through it thot uses a Constructive an destrudive intesferenceLight Source | : The miaoscope uses 0 bright igh source , typrcaly a Halogen or LED Lamp to illuminole the specimen Annular Diaphragm 2H fs looted ih the condenser and adjust the Phase Plate light 40 come feom all lrections + Ti is lomted in objective lens, The phase plate inroduea a phase shift to the light thot passes through It that causes a Constructive and destrudive intesfesence Tnage Fermation |: The interference between light waves results in diferences in brightness € @nhst In Fal image This makes the details of sampk more visible* The interference between light waves results in diferences in brightness - @nhaat (n Final image This makes the details of sampk more visible Tmmage Formation APPLICATION e Tt helps to see details in ting. Cells without staining. them © T is used for visualisation of vosous cell organdles Ike Milochondlia Nucleus etc eT is used to examine qils in medical samples fos abnosmalities © Tt allows obsenvalion of Bacteria Other tiny. Microorganisms AADVANTAGES © No need for stoinin © Provides high conkrost images oF cells © st Effedive © Usefiu for study of living, ells -€ dynamic processes OisapvaNTAGeES © D generally offer, low resolution compared to some other Microscopes ike electron microscope © TF may not work well with very thick -@ dense specimenDark Fieto Microscope © A Dak Field Microscope is 0 type of Microsmpe thot makes it ensy to see tiny or transparent objects © TF adjust light on sample in auch a woy thal object ise appears brignt while the eee wil be dark, almost black, © This technique highligh delaiis that might be hard fo see with regular microscopes.Feqular microscopes PRINCIPLE The principle of Dak Field Microscope involves creating, G br image of specimen againat davk background by. Using, angled light through an opaque disk & Eyepiece LensEyepiene. Leng Objective Lens Unrefracted Re ‘ Specimen Condenser Opaque Disk ey 5@s— Light Source ansKey Components € Working MecHanism + TF poovides ilurnination typically 0 bright focused! beam of light Dork Field Condenser]: Tt iS @ gpecial Gondenser consist OF an Opaque Disk 4hat blocks direct light from entesing. the Objective lens Satiesing, |: When the sampe is illuminated . tt scatters the light This scattered light now coming, fom different angles is colleded by objective leng @ | Light _Sowce© Lugar sore j+ it provides muminaton typamy G Dngnr Tocusect beam of light Dark Field Gondenses |: Tt is @ special Gndenser consist OF an Opaque. Disk that blacks direct light from entesing. the objective lens @ | Saottesing, |: When the samde fs illuminated . it scatters +he light: This scoltered [ight now coming, foom different angles is collected by objective lena. “mage Formation]: The objective lens focuses this scltered light fo form a Daght image Gf specimen against a das backysoundfom a bript image. of specimen again a ask bactsound . APPLICATION © F haps scientist see live cla € baceia without needing, to add dyes oF is usd to view vey small pastides like nanopantices to check their size € shope. © TF allows doctosa to see fle detoila in tise samples that are hard to spot with eqular tethniquerADVANTAGES. © F ehances contest of Image © Not fequire! stainin °F is effective for visualsing, very fine detail, DisabvanTAGeSs © The ovesall image may appeas dimmer compased to eight fie microscopy beause it velies on scoltesed light alhes: than direct lightEvectron Microscope © An Electron Mictoscope is a microscope that uses a beam of acelerated electrons instead of visible light a8 a xouce of ilumination © The Wavelength of dlectrona can be upto 100,000 times shostes than that oF Visible light - © TF Is of two types © Transmission Electron Microscope @ Scanning. Becton MicroscopeTransmission Electron Microscope is a high “wesolution Imaging. technique that uses beam of eledrons, 4ranamitted through on ultra thin Specimen to produce detailed images Principe Tranamission Eledron Microscopy operates on the painciple of ‘renaming a beam of aedrong ‘through a very thin specimen ond foomin: an image based on +he interactiong between the electona ani tho anmaloWorking MecaNisH The working, mechanism of Transmission Becton Microstopy involves Several key steps © Sample Pseparation © Electron Source CElecton Gun ) © Beam Tranamission © Tage Formation @ Tmage Detection ° mage Analysis© mage Detection Tinage Analysis —AppucAiON © Transmission Elechon Microscopy is used to obsesue fihe shuchim of matesiols at nanosale tnduding the study of crystal Strucures , defects and interfaces °F assist in Ghagnosing diserse by examining Obnosmalities lulay — ShuchiresScanning Evectron Microscopy Scanning, Electron Microscopy i a powerful te technique used fo bsewe the surface stricture of samples” at high magnification Working MecHanism The working of scanning, electron microscopy involves Severa) ey steps © Sample Feeparation © Electron Gun ag ledron SowreMagnification Working Mecianism The working of scanning, electron micrascopy thuclues several key steps ss © Sample Foeparation ledron Gun a& Bledron Sowre © Beam Focusing, © Ganning © Therion With Sample © Signa Detection © Tage formationPrincipe. The painciple of Scanning, electron microscopy Involves scannin focused beam of elecrong across the surface of a somple and detecting, the emitted signals to creole on mage AppLcATION © for analysis of surfoxe features, € (ughness of moteniala like metal , polymers € @ramide . © Examination of pothalogical amples ach a& fiscues € biopsies to identify disease (elated changes .