1、Designation: E211 82 (Reapproved 2016)Standard Specification forCover Glasses and Glass Slides for Use in Microscopy1This standard is issued under the fixed designation E211; the number immediately following the designation indicates the year oforiginal adoption or, in the case of revision, the year
2、 of last revision. A number in parentheses indicates the year of last reapproval. Asuperscript epsilon () indicates an editorial change since the last revision or reapproval.1. Scope1.1 This specification describes glass covers and slides foruse in routine microscopy.1.2 The values stated in SI unit
3、s are to be regarded asstandard. No other units of measurement are included in thisstandard.2. Glass2.1 Cover GlassGlass shall be colorless, transparent glass,sufficiently free of pits, bubbles, or cloudiness so as not tointerfere with end-use performance. Such defects shall beregarded as major defe
4、cts and limited accordingly. Inspectionshall be by the unaided eye in a good light viewing a singlecover glass at a time. Refractive index at sodium D line (589.3nm) shall be 1.523 6 0.005.2.2 Glass SlidesGlass slides covered by this specificationshall consist of colorless, transparent glass which m
5、ay have aslight greenish tint when viewed edgewise. It shall be suffi-ciently free of pits, nicks, bubbles, striae, scratches, andcloudiness when observed by the unaided eye so as not toimpair end-use performance. Index of refraction at the sodiumD line (589.3 nm) shall be 1.50 to 1.54.COVER GLASSES
6、3. Dimensions3.1 General UseThe dimensions of cover glass for gen-eral use shall be as follows:3.1.1 Square22 mm on a side.3.1.2 Rectangular22 by 30, 22 by 40, 22 by 50, or 22 by60 mm and 24 by 30, 24 by 40, 24 by 50, and 24 by 60 mm.3.1.3 ThicknessThe thickness shall be as follows:Number Thickness,
7、 mm1 0.13 to 0.17112A0.16 to 0.192 0.17 to 0.25ASpecified as optimum by most microscope manufacturers.3.2 Limited UseThe following sizes while recognized,find limited use and are generally not as readily available.3.2.1 Square18, 24 and 25 mm on a side.3.2.2 Rectangular24 by 55 mm, 35 by 50 mm, 45 b
8、y 50mm, 20 by 26 mm and 20 by 40 mm.3.2.3 Circular18, 22 or 25 mm in diameter.3.2.4 ThicknessThickness Number 0, 0.08 to 0.13 mm.3.3 ToleranceTolerance on dimensions other than thick-ness shall be 60.5 mm.3.4 Blood Counting ChambersThe dimensions of coverglasses for blood counting chambers shall be
9、as follows:Length, mmAwidth, mmAThickness, mmB20 21 0.4 or 0.620 26 0.4 or 0.625 28 0.4 or 0.6AThe tolerance on length and width shall be 1.0 mm.BThe tolerance on thickness shall be 0.06 mm.4. Planeness4.1 General UseCover glass may be expected to pass thetest as described in 14.1.4.2 Blood Counting
10、 ChambersCover glass shall be planewithin 0.002 mm on both sides and shall pass the planeness testas described in 16.2.5. Corrosion Resistance5.1 While no corrosion tests for cover glasses are herebyrequired, when desired for critical use, use the test in 14.2.GLASS SLIDES6. Dimensions6.1 General Us
11、e:6.1.1 Size75.5 by 25.5 6 0.8 mm.6.1.2 Thickness A1.0 6 0.15 mm.Thickness B1.2 6 0.15 mm.6.2 Darkfield and Interference Microscopy:1This specification is under the jurisdiction of ASTM Committee E41 onLaboratory Apparatus and is the direct responsibility of Subcommittee E41.01 onLaboratory Ware and
12、 Supplies.Current edition approved Sept. 1, 2016. Published September 2016. Originallyapproved in 1963. Last previous edition approved in 2010 as E211 82 (2010).DOI: 10.1520/E0211-82R16.Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States1
13、6.2.1 Size75.5 by 25.5 6 0.8 mm.6.2.2 Thickness C1.20 6 0.005 mm.Thickness D0.90 6 0.075 mm.Thickness E1.50 6 0.05 mm.6.3 Embryology37.5 6 0.8 or 50.5 6 0.8 by 75.5 6 0.8mm.6.3.1 ThicknessSame as either thickness A or thickness Bin 6.1 on general-use glass slides.6.4 Petrography:6.4.1 Size25.5 by 45
14、.5, 27 by 46, or 28 by 48 6 0.8 mm.6.4.2 ThicknessSame as thickness A or B under general-use slides.7. Planeness and Parallelism7.1 General UseNo requirements.7.2 Darkfield, Interference, Embryology, PetrographyNorequirements.8. Corrosion Resistance8.1 General Use and Darkfield and InterferenceMicro
15、scopyThe slides shall not be adversely affected byboilingnor shall they exhibit any pink color in the solubilitytest.8.2 Embryology, PetrographyNo requirements.9. Workmanship9.1 All edges of all glass slides shall be polished or ground.They shall pass the wettability tests in Section 17. General-use
16、microscope slides shall be smooth and free from nicks orblemishes on the short edge which might impair their service-ability for blood smear work.9.2 Frosted EndOne end of one or both surfaces ofmicroscope slides may be frosted to allow marking with a No.2 pencil or ballpoint pen for a distance of 1
17、9 6 3 mm from theend. Alternatively, both ends of the microscope slide may befrosted for a distance of 9 6 2 mm.GENERAL10. Sampling10.1 The cover glass and glass slides to be tested forcompliance with these specifications shall be selected atrandom from the lot or shipment. Ten or more cover glasses
18、 orslides shall be tested for index of refraction and clarity,resistance to boiling, and solubility.11. Packaging and Package Marking11.1 Cover GlassGeneral-use cover glass shall be pack-aged in lint-free boxes generally standing on edge so that theglass is convenient to use without contamination. I
19、t should beprotected from infiltrating dust and moisture which can causesticking together of the glass.11.2 Glass SlidesSlides shall generally stand on edge inunit packages to present them conveniently for use by the enduser without contamination or loss of cleanliness. Packagesshall be sufficiently
20、 lint-free to avoid contamination. Unitpackages may be wrapped in a moisture-proof barrier material,either singly or in groups to reduce field contamination,sticking and to extend shelf life.TEST METHODS12. Index of Refraction12.1 Measure with a refractometer and sodium (yellow)light.13. Clarity13.1
21、 Examine the defects by the unaided eye. This meansthe use of an emmetropic eye or an ametropic eye corrected tonormal vision without further magnification. Examine coverglasses in good light in front of a dark surface. Examinegeneral-use slides with daylight or diffuse uniform artificiallight again
22、st a uniform dark background. Examine darkfieldand interference microscopy slides with a 10 magnifier in alight beam of at least 25 fc against a black background.14. Planeness14.1 The number of cover glasses specified in Table 1 shallbe stacked on a flat surface. The height of the stack shall bemeas
23、ured within 60.05 mm. A thin stiff aluminum plate cut tothe size of the cover glass is to be placed on top of the stack.A weight is placed on top of the plate so that the total mass ofthe aluminum plate plus the weight is 100 g. Again, the heightof the stack is to be measured to within 6 0.05 mm. Th
24、edifference between the unloaded and loaded readings is thecompression. Compression shall not exceed the amount shownin the Table 1.14.2 Test planeness against an optical flat, using the inter-ferometric method described in National Bureau of StandardCircular 507.15. Thickness Variation15.1 Permissi
25、ble thickness variation in any one specimen ofcover glass is as follows:15.1.1 For sizes up to and including 25 mm in any onedimension, not more than 1 % of the specimens measured shallshow a greater variation than 0.015 mm in any one specimen.15.1.2 For sizes above 25 mm in any one dimension, notmo
26、re than 1 % of the specimens measured shall show a greatervariation than 0.020 mm in any one specimen.16. Resistance to Corrosion and Other Defects16.1 Resistance to BoilingImmerse the slides in distilledwater in a closed vessel having a small steam vent and boilcontinuously for 6 h, occasionally ad
27、ding boiling water toTABLE 1 Quantity of Cover Glasses and Compression Allowed(22 by 22-mm Cover Glass)ThicknessNo.Quantity ofCover GlassCompression(Warpage), mm0 140 2.131 100 1.42112 86 1.192 70 1.01E211 82 (2016)2compensate for evaporation losses. Suspend the slides in waterso that they do not co
28、me in contact with the containing vessel.The slides shall show no evidence of fogging, chipping, orcracking as a result of the test.16.2 Solubility of Cover GlassesClean 24 cover glasses byimmersion for 1 min in distilled water, remove, and air-dry for1 min in a vertical position. Perform this clean
29、ing operationthree times, using a new quantity of distilled water for eachimmersion. Half fill an alkali-resistant 750-mL Erlenmeyerflask with distilled water and boil for 10 min. Allow the waterto cool for 3 min and then decant until the volume of the flaskis approximately 100 mL. Keeping the flask
30、 covered, let itstand until the water has cooled to approximately 70C. Add0.2 mL of USP standard phenolphthalein solution and continueto cool to 60C. Add the cover glasses, one at a time.Superimpose the cover glasses by tilting and gently swirlingthe flask. Let the solution cool toward room temperat
31、urewithout agitation for 1 h. Absorption of CO2from the atmo-sphere must be avoided during the test. No pink color shall bevisible through the 24 superimposed cover glasses.16.3 Solubility of Glass SlidesClean each individual slideby immersing for 1 min in distilled water, removing, and airdrying fo
32、r 1 min in a vertical position. Perform this cleaningoperation three times, using a new quantity of distilled waterfor each immersion. After the final cleaning operation, placethe slides immediately in distilled water containing 0.2 % USPstandard phenolphthalein solution which has just been boiledfo
33、r 10 min in a flask (which in itself will not affect the testsolution) and allow to cool (preferably without excessiveagitation but with mild stirring with a thermometer) to 60C.Place the slides in the flask so as to obtain the maximumexposure of area per slide. No pink color shall develop in thetes
34、t solution at any time during the ensuing 1 h, the solutioncooling toward room temperature at that time. Absorption ofCO2from the atmosphere must be avoided during the 1-hstanding.17. Wettability17.1 Micro SlidesTo assure that blood or other test mate-rial will spread evenly across the surface of th
35、e slide, thesurface of the slide should not be touched, handled, orfingerprinted on or near the edges and surfaces to be used. Thesample slide shall be dipped not more than half way into abeaker of distilled water, and then examined for properdispersion of the liquid and absence of gaps in the film
36、oneither side of the slide.17.2 Cover GlassIf wettability is critical, the followingtest should be used. Grasp cover glass by edges near one end.Dip bottom half into distilled water. Upon removing, coverglass should show an even coat of water on both sides with nodry spots or beading. If some beadin
37、g does occur, measure thecontact angle between the horizontal plane of the cover glassand the drop of distilled water. Reject cover glass if contactangle is greater than 30.18. Packaging and Package Marking18.1 Product DescriptionPackages for both slides andcover glasses should include size, thickne
38、ss, special informa-tion such as selected, frosted end or precleaned, stock numberif applicable, quantity or weight, name of manufacturer ordistributor, and country of origin. Lot numbers or othermanufacturing data shall appear on either the unit or over-packer.18.2 Packages of slides or cover glass
39、es meeting thesespecifications may be labeled “Meets ASTM SpecificationE211, Section _ .”19. Keywords19.1 cover glass; glass; microscopy; slidesASTM International takes no position respecting the validity of any patent rights asserted in connection with any item mentionedin this standard. Users of t
40、his standard are expressly advised that determination of the validity of any such patent rights, and the riskof infringement of such rights, are entirely their own responsibility.This standard is subject to revision at any time by the responsible technical committee and must be reviewed every five y
41、ears andif not revised, either reapproved or withdrawn. Your comments are invited either for revision of this standard or for additional standardsand should be addressed to ASTM International Headquarters. Your comments will receive careful consideration at a meeting of theresponsible technical comm
42、ittee, which you may attend. If you feel that your comments have not received a fair hearing you shouldmake your views known to the ASTM Committee on Standards, at the address shown below.This standard is copyrighted by ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19
43、428-2959,United States. Individual reprints (single or multiple copies) of this standard may be obtained by contacting ASTM at the aboveaddress or at 610-832-9585 (phone), 610-832-9555 (fax), or serviceastm.org (e-mail); or through the ASTM website(www.astm.org). Permission rights to photocopy the standard may also be secured from the Copyright Clearance Center, 222Rosewood Drive, Danvers, MA 01923, Tel: (978) 646-2600; http:/ 82 (2016)3
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