1、BSI Standards PublicationBS ISO 8036:2015Microscopes Immersionliquids for light microscopyBS ISO 8036:2015 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of ISO 8036:2015. It supersedes BS ISO 8036:2006 which is withdrawn.The UK participation in its preparation was e
2、ntrusted to Technical Committee CPW/172, Optics and Photonics.A list of organizations represented on this committee can be obtained on request to its secretary.This publication does not purport to include all the necessary provisions of a contract. Users are responsible for its correct application.
3、The British Standards Institution 2015.Published by BSI Standards Limited 2015ISBN 978 0 580 88295 1 ICS 37.020 Compliance with a British Standard cannot confer immunity from legal obligations.This British Standard was published under the authority of the Standards Policy and Strategy Committee on 3
4、0 June 2015.Amendments/corrigenda issued since publicationDate T e x t a f f e c t e d ISO 2015Microscopes Immersion liquids for light microscopyMicroscopes Liquides dimmersion pour microscopie optiqueINTERNATIONAL STANDARDISO 8036Second edition 2015-06-01Reference number ISO 8036:2015(E)BS ISO 8036
5、:2015ISO 8036:2015(E)ii ISO 2015 All rights reservedCOPYRIGHT PROTECTED DOCUMENT ISO 2015, Published in SwitzerlandAll rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including pho
6、tocopying, or posting on the internet or an intranet, without prior written permission. Permission can be requested from either ISO at the address below or ISOs member body in the country of the requester.ISO copyright officeCh. de Blandonnet 8 CP 401CH-1214 Vernier, Geneva, SwitzerlandTel. +41 22 7
7、49 01 11Fax +41 22 749 09 47copyrightiso.orgwww.iso.orgBS ISO 8036:2015ISO 8036:2015(E)Foreword iv1 Scope . 12 Normative references 13 Classification 14 Characteristics of immersion liquids 14.1 Optical properties . 14.2 Transmittance 24.3 Autofluorescence. 24.4 Viscosity 24.5 Density . 25 Minimum r
8、equirements 36 Ingredients . 37 Designation of immersion liquids 38 Labelling of containers. 39 Technical data sheet 4 ISO 2015 All rights reserved iiiContents PageBS ISO 8036:2015ISO 8036:2015(E)ForewordISO (the International Organization for Standardization) is a worldwide federation of national s
9、tandards bodies (ISO member bodies). The work of preparing International Standards is normally carried out through ISO technical committees. Each member body interested in a subject for which a technical committee has been established has the right to be represented on that committee. International
10、organizations, governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.The procedures used to develop this document and those intended for
11、its further maintenance are described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the different types of ISO documents should be noted. This document was drafted in accordance with the editorial rules of the ISO/IEC Directives, Part 2 (see www.iso.org/
12、directives).Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of any patent rights identified during the development of the document will be in
13、 the Introduction and/or on the ISO list of patent declarations received (see www.iso.org/patents).Any trade name used in this document is information given for the convenience of users and does not constitute an endorsement.For an explanation on the meaning of ISO specific terms and expressions rel
14、ated to conformity assessment, as well as information about ISOs adherence to the WTO principles in the Technical Barriers to Trade (TBT), see the following URL: Foreword Supplementary information .The committee responsible for this document is ISO/TC 172, Optics and photonics, SC 5, Microscopes and
15、 endoscopes.This second edition cancels and replaces the first edition (ISO 8036:2006), where in Clause 6, the outdated reference to the EU Directive 91/155/EEC and its Amendments 93/112/EC and 01/58/EC were removed.iv ISO 2015 All rights reservedBS ISO 8036:2015Microscopes Immersion liquids for lig
16、ht microscopy1 ScopeThis International Standard describes the characteristics of immersion liquids used in microscopy. It classifies immersion liquids according to their field of application and specifies requirements and test methods for each type.This International Standard further specifies a sys
17、tem of designation for immersion liquids, the information to be included on container labels, and the information to be supplied in technical data sheets.2 Normative referencesThe following documents, in whole or in part, are normatively referenced in this document and are indispensable for its appl
18、ication. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies.ISO 2592, Determination of flash and fire points Cleveland open cup methodISO 8255-1, Microscopes Cover glasses Part 1: Dimensional
19、tolerances, thickness and optical propertiesISO 8255-2, Microscopes Cover glasses Part 2: Quality of materials, standards of finish and mode of packaging3 ClassificationDepending on their field of application, immersion liquids are classified as follows: type N: immersion oil for general use in ligh
20、t microscopy; type F: immersion oil which meets the requirements of fluorescence microscopy; type G: spectrally pure glycerol (commonly known as glycerine) for glycerol immersion.4 Characteristics of immersion liquids4.1 Optical propertiesThe optical properties of immersion liquids are defined by th
21、e refractive index at the wavelength = 546,07 nm, ne, at a defined temperature (23 C) and pressure (1 013,25 hPa), as well as by the Abbe number (reciprocal of the dispersive power), ve.The Abbe number, ve, is calculated using Formula (1):eeFC=nnn1(1)wherenFis the refractive index at = 479,99 nm;nCi
22、s the refractive index at = 643,85 nm.INTERNATIONAL STANDARD ISO 8036:2015(E) ISO 2015 All rights reserved 1BS ISO 8036:2015ISO 8036:2015(E)The refractive indices are usually measured with an Abbe refractometer at constant temperature with a tolerance of 0,1 C. A mercury-cadmium spectral lamp is use
23、d as the light source.4.2 TransmittanceModern detection methods in light microscopy require an extension of the spectral range of immersion liquids.The transmittance is measured using a suitable spectrophotometer in quartz cuvettes with an optical path length through the immersion liquid of d = 10 m
24、m against an empty reference cuvette.4.3 AutofluorescenceThe autofluorescence of immersion liquids is measured with a fluorescence spectrometer. Quinine sulfate in 0,05 mol/l sulfuric acid is used as the fluorescence standard. For the fluorescence measurement, the following excitation and emission w
25、avelengths are used: F(365 nm/450 nm) = fluorescence excitation at 365 nm, measurement of fluorescence emission at 450 nm; F(405 nm/485 nm) = fluorescence excitation at 405 nm, measurement of fluorescence emission at 485 nm.As the fluorescence emission decreases during the measurement, the autofluor
26、escence of immersion liquids is averaged over a measurement time of 60 s.Immersion liquid samples and standard liquids are measured in a fluorescence cuvette made of quartz or special optical glass with an optical path length through the immersion liquid of d = 10 mm. The reagents used shall meet th
27、e quality standards for fluorescence spectroscopy.Prepare a quinine sulfate stock solution of 500 mg/l with 0,05 mol/l sulfuric acid. Before use, prepare two standard solutions of 0,1 mg/l and 2,5 mg/l quinine sulfate in 0,05 mol/l sulfuric acid by diluting the stock solution 1:5000 and 1:200, respe
28、ctively. Store the stock solution in a closed brown glass bottle at 6 C to 12 C for no more than two months. Always prepare the standard solutions fresh on the day of use.As an alternative to quinine sulfate solution, solid-state fluorescence standards with the same shape as a 10 mm cuvette may be u
29、sed. These shall be calibrated against a quinine sulfate standard solution at least once a year.NOTE 1 Solid-state fluorescence standards are available as spectroscopy accessories.The information regarding the autofluorescence of the immersion liquids is given in terms of the equivalent amount of qu
30、inine sulfate, in mg/l.NOTE 2 Quinine sulfate is more suitable for use as a fluorescence standard than rhodamine B because immersion oils tend, like quinine sulfate, to emit in the blue region in the case of UVA excitation, while rhodamine B emits in the green region.4.4 ViscosityThe viscosity of th
31、e immersion liquid shall be measured by means of a capillary viscometer, e.g. an Ubbelohde viscometer, at a temperature of 23 C.4.5 DensityThe density of the immersion liquid shall be measured at a temperature of 23 C by means of a hydrometer or a pyknometer.2 ISO 2015 All rights reservedBS ISO 8036
32、:2015ISO 8036:2015(E)5 Minimum requirementsSee Table 1.6 IngredientsImmersion liquids conforming to this International Standard shall not contain ingredients which are known to have any adverse effects on safety or health.7 Designation of immersion liquidsImmersion liquids conforming to this Interna
33、tional Standard shall be designated as in the following example:EXAMPLE Immersion liquid ISO 8036 Type F.8 Labelling of containersImmersion liquids are commonly packed and marketed in brown glass bottles or in oilers made from polyethylene or polypropylene. The labelling of these containers shall in
34、clude the following information: the product name; the designation in accordance with this International Standard; labelling required by regulations for hazardous freight; the refractive index, ne, at 23 C; the Abbe number, ve; the batch number; the expiration date when stored in the unopened origin
35、al packaging in accordance with the manufacturers instructions; information on how to obtain technical and safety data; the name and address of the manufacturer and the supplier, plus the country of origin if different from that given in the manufacturers address.Table 1 Minimum requirements for imm
36、ersion liquidsaParameter Type N (standard)Type F (for fluorescence)Type G (glycerol)bReference temperature (C)Tolerance (C)230,1230,1230,1Spectral range (nm) 400 to 900 320 to 1 100 230 to 1 100Refractive index, ne1,518 0 0,000 5 1,518 0 0,000 5 1,450 0,005Abbe number, ve43 4 43 4 58 4Transmittance
37、(d = 10 mm)at 1 100 nm 80 % 70 %aImmersion liquids, with the exception of glycerol, may not contain volatile or hygroscopic ingredients.Specifications for these liquids assume the use of a cover glass as specified in ISO 8255-1 and ISO 8255-2.bGlycerol is hygroscopic and its physical properties depe
38、nd on its water content.cAutofluorescence of type N oil has no specified values, but high values can be expected. ISO 2015 All rights reserved 3BS ISO 8036:2015ISO 8036:2015(E)Parameter Type N (standard)Type F (for fluorescence)Type G (glycerol)bat 1 000 nm 80 % 70 %at 900 nm 90 % 90 % 90 %at 800 nm
39、 95 % 95 % 98 %at 760 nm 95 % 95 % 98 %at 600 nm 95 % 95 % 98 %at 500 nm 95 % 95 % 98 %at 450 nm 85 % 95 % 98 %at 420 nm 75 % 95 % 98 %at 400 nm 60 % 95 % 98 %at 380 nm 90 % 95 %at 365 nm 80 % 95 %at 350 nm 70 % 90 %at 330 nm 40 % 80 %at 320 nm 20 % 70 %at 310 nm 50 %at 230 nm to 300 nm 30 %Autofluo
40、rescence (mg/l quinine sulfate)F(365 nm/450 nm)c0,06 0,02F(405 nm/485 nm)c1,20 0,50F(313 nm/450 nm) 0,05Viscosity (mm2/s)at 23 C 0,1 C 50 to 1 500 50 to 1 500baImmersion liquids, with the exception of glycerol, may not contain volatile or hygroscopic ingredients.Specifications for these liquids assu
41、me the use of a cover glass as specified in ISO 8255-1 and ISO 8255-2.bGlycerol is hygroscopic and its physical properties depend on its water content.cAutofluorescence of type N oil has no specified values, but high values can be expected.9 Technical data sheetThe manufacturer or the supplier of an
42、 immersion liquid shall make a technical data sheet available on request. This data sheet shall contain the following information: product name; manufacturer; designation in accordance with this International Standard; field of application; chemical basis; reference temperature for refractive index
43、measurements; spectral range;Table 1 (continued)4 ISO 2015 All rights reservedBS ISO 8036:2015ISO 8036:2015(E) refractive indices ne, nD(at sodium D-line), ng(at mercury g-line), nF, nC; Abbe number, ve; temperature coefficient of refractive index in the range 15 C to 40 C; transmittance within the
44、spectral range for an optical path length d = 10 mm; autofluorescence; behaviour with regard to microscopy stains; density at 23 C; viscosity at 23 C; flash point determined in accordance with ISO 2592 (Cleveland open cup method); storage conditions; recommended cleaning agents; information on dispo
45、sal. ISO 2015 All rights reserved 5BS ISO 8036:2015ISO 8036:2015(E) ISO 2015 All rights reservedICS 37.020Price based on 5 pagesThis page deliberately left blankBSI is the national body responsible for preparing British Standards and other standards-related publications, information and services.BSI
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