Cryocell The Cryocell Sperm counting chamber is a simple to use device for rapid and accurate Sperm count & motility evaluation directly from undiluted Semen. 10 MICRON DEPTH DESCRIPTION The Chamber is composed of two parts: The lower main part has a glass base wherein the centre portion has been coated with a metal layer on which there is grid of 0.01 sq. mm. This part has been lowered by 10 microns as compared to the rest of the chamber. It is on this portion that a very small drop of the sample is placed. Figure: The upper part is cover glass, which is to placed on the drop of the sample & firmly pressed on both the sides to remove the excessive sample, thereby ensuring that the space between the cover slip & the Grid is exactly 10 microns. PREPRATION OF THE CHAMBER Before use, make certain that the opposed glass surfaces are absolutely clean and free of dust, since the size of the particles is more than 10 microns. You should brush the surfaces with a very soft brush.(Provided with the pack) This is a high precision instrument built to very narrow tolerances. Thus extra care & cleanliness is advised. The depth of the chamber is guaranteed at 10.0 microns ( + 0.001 microns). Any speck of dust deposited on each side of the chamber or in chamber itself can render this accuracy void. Please therefore carefully clean all surfaces of the chamber & the cover slip with water and a brush & thereafter with special Lens paper & a dry brush.
CryocellThe Cryocell Sperm counting chamber is a simple to use
device for rapid and accurate Sperm count & motility
evaluationdirectlyfromundilutedSemen.10MICRONDEPTHDESCRIPTIONTheChamberiscomposedoftwoparts:Thelowermainparthasaglassbasewhereinthecentreportionhasbeencoatedwithametallayeronwhichthereisgridof0.01sq.mm.Thisparthasbeenloweredby10micronsascomparedtotherestofthechamber.Itisonthisportionthataverysmalldropofthesampleisplaced.Figure:Theupperpartiscoverglass,whichistoplacedonthedropofthesample&firmlypressedonboththesidestoremovetheexcessivesample,therebyensuringthatthespacebetweenthecoverslip&theGridisexactly10microns.PREPRATIONOFTHECHAMBERBeforeuse,makecertainthattheopposedglasssurfacesareabsolutelycleanandfreeofdust,sincethesizeoftheparticlesismorethan10microns.Youshouldbrushthesurfaceswithaverysoftbrush.(Providedwiththepack)Thisisahighprecisioninstrumentbuilttoverynarrowtolerances.Thusextracare&cleanlinessisadvised.Thedepthofthechamberisguaranteedat10.0microns(+0.001microns).Anyspeckofdustdepositedoneachsideofthechamberorinchamberitselfcanrenderthisaccuracyvoid.Pleasethereforecarefullycleanallsurfacesofthechamber&thecoverslipwithwaterandabrush&thereafterwithspecialLenspaper&adrybrush.SEMENANAYSISMixthespecimenwell,takingcaretoavoidformationofbubbles,withtheaidofarod,putasmalldropinthecentreofthecentrestrip.Graspthecoverglass&Placeitontheedges&pressfirmly.Thisensurestheautomaticspreadofthedropintothethicknessof10microns.Itisrecommendedthatthedropshouldspreadontheentireareaofthecentreportion.Oncethecoverglassisinplaceavoidfurthertouching.Liftingandrecoveringasthismayaffecttheuniformspreadofthespermwithinthedrop.Liftthechamber&placeisonthestageofthemicroscopeusinga20xobjectiveanda10xeyepiece.Avoidusingthe10xobjective,asthespermswithetoosmalltosee.You
can use a 40x objective to do the morphological examination as well
which was not possible with the
conventionalchambersduetothethicknessoftheircoverglasses.Afterfocusingmovethestageofthemicroscopeandlocatethegridinthecentreoftheviewarea.UnevendistributionoftheSpermsmeanthatthesamplewasnotmixedthoroughly.Ifspermscannotbeseeninonefocalplane,impropercleaningoftheglasssurfacesissuspected.Ineithercaserepeatthisprocedureaftercorrectinghefault.MOTILITYESTIMATIONCountallthenonmotilespermswithin(10continuoussquareseitherhorizontallyorvertically).Thencountthemotilespermsinthesamearea.Addingupboththenumberswouldgiveyouthetotalcountperml.(inMillions).Youcanthendividethemotilewiththetotalcounttoreachthepercentagemotility.Usingthesamemethodestimatethegradeofthemotilityfrom+1to+4.Thisestimationismuchmoreaccuratethanthatperformedfromtheordinaryslideswherethespermsmaybecompressedbythecoverslipandtheirmovementimpaired.Thecalculationthiswayismuchbetterthanonedoneonahemoctometerwhereoneendsuponlygettingthetotalcount¬themotility.SPECIALCASESBubbles:Ifbubblesarepresentinthegridarea,itisrecommendedthatthedropmaybereplacedbyanother,unlessthebubblesaresmallenoughnottointerferewiththeanalysis.Largeparticlesofdust,threadsetc.willalsointerferewiththecountbychangingthedepthofthespace&insuchacasethedropshouldbereplaced.Majorvariationincountsbetweendropsofthesamespecimenmeans,thatthesamplewasnotmixeswelloristooviscoustomixcompletely.CLEANINGANDPREPRATIONFORREUSEDipthebrushintocleanwaterandsimplywipebothsidesoftheglasses.Thensqueezethebrushandspongetheremainingwateroffthechamber.Finallydrythesurfaceswithspeciallenspaper.(Providedwiththepack)TheChamberisnowreadyforreuse.
