1、BSI Standards Publication BS ISO 8036:2015 Microscopes Immersion liquids for light microscopyBS ISO 8036:2015 BRITISH STANDARD National foreword This 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 It supersedes BS ISO 8036:2006 which is withdrawn.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
2、 was entrusted 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 appli
3、cation. The British Standards Institution 2015. Published by BSI Standards Limited 2015 ISBN 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 Com
4、mittee on 30 June 2015. Amendments/corrigenda issued since publication Date T e x t a f f e c t e d ISO 2015 Microscopes Immersion liquids for light microscopy Microscopes Liquides dimmersion pour microscopie optique INTERNATIONAL STANDARD ISO 8036 Second edition 2015-06-01 Reference number ISO 8036
5、:2015(E)BS ISO 8036:2015ISO 8036:2015(E)ii ISO 2015 All rights reserved COPYRIGHT PROTECTED DOCUMENT ISO 2015, Published in Switzerland All 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 mech
6、anical, including photocopying, 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 office Ch. de Blandonnet 8 CP 401 CH-1214 Vernier, Geneva,
7、Switzerland Tel. +41 22 749 01 11 Fax +41 22 749 09 47 copyrightiso.org www.iso.orgBS ISO 8036:2015ISO 8036:2015(E)Foreword iv 1 Scope . 1 2 Normative references 1 3 Classification 1 4 Characteristics of immersion liquids 1 4.1 Optical properties . 1 4.2 Transmittance 2 4.3 Autofluorescence. 2 4.4 V
8、iscosity 2 4.5 Density . 2 5 Minimum requirements 3 6 Ingredients . 3 7 Designation of immersion liquids 3 8 Labelling of containers. 3 9 Technical data sheet 4 ISO 2015 All rights reserved iii Contents PageBS ISO 8036:2015ISO 8036:2015(E) Foreword ISO (the International Organization for Standardiza
9、tion) is a worldwide federation of national standards 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
10、represented on that committee. International 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
11、 develop this document and those intended for 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 th
12、e ISO/IEC Directives, Part 2 (see www.iso.org/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 du
13、ring the development of the document will be in 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
14、meaning of ISO specific terms and expressions related 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
15、 172, Optics and photonics, SC 5, Microscopes and 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
16、ISO 8036:2015Microscopes Immersion liquids for light microscopy 1 Scope This 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
17、. This International Standard further specifies a system 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 references The following documents, in whole or in part, are normatively referen
18、ced in this document and are indispensable for its application. 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 method I
19、SO 8255-1, Microscopes Cover glasses Part 1: Dimensional tolerances, thickness and optical properties ISO 8255-2, Microscopes Cover glasses Part 2: Quality of materials, standards of finish and mode of packaging 3 Classific ation Depending on their field of application, immersion liquids are classif
20、ied as follows: type N: immersion oil for general use in light 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 liquids 4.1 Optical properti
21、es The optical properties of immersion liquids are defined by the refractive index at the wavelength = 546,07 nm, n e , at a defined temperature (23 C) and pressure (1 013,25 hPa), as well as by the Abbe number (reciprocal of the dispersive power), v e . The Abbe number, v e , is calculated using Fo
22、rmula (1): n nn (1) where n F is the refractive index at = 479,99 nm; n C is the refractive index at = 643,85 nm. INTERNATIONAL ST ANDARD 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
23、 temperature with a tolerance of 0,1 C. A mercury-cadmium spectral lamp is used as the light source. 4.2 Transmittance Modern detection methods in light microscopy require an extension of the spectral range of immersion liquids. The transmittance is measured using a suitable spectrophotometer in qua
24、rtz cuvettes with an optical path length through the immersion liquid of d = 10 mm against an empty reference cuvette. 4.3 A ut ofluor esc enc e The autofluorescence of immersion liquids is measured with a fluorescence spectrometer. Quinine sulfate in 0,05 mol/l sulfuric acid is used as the fluoresc
25、ence standard. For the fluorescence measurement, the following excitation and emission wavelengths 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 emissio
26、n at 485 nm. As the fluorescence emission decreases during the measurement, the autofluorescence 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 opti
27、cal path length through the immersion liquid of d = 10 mm. The reagents used shall meet the 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
28、 sulfate in 0,05 mol/l sulfuric acid by diluting the stock solution 1:5000 and 1:200, respectively. 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
29、solution, solid-state fluorescence standards with the same shape as a 10 mm cuvette may be used. 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
30、the autofluorescence of the immersion liquids is given in terms of the equivalent amount of quinine 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
31、of UVA excitation, while rhodamine B emits in the green region. 4.4 Viscosity The viscosity of the immersion liquid shall be measured by means of a capillary viscometer, e.g. an Ubbelohde viscometer, at a temperature of 23 C. 4.5 Density The density of the immersion liquid shall be measured at a tem
32、perature of 23 C by means of a hydrometer or a pyknometer.2 ISO 2015 All rights reservedBS ISO 8036:2015ISO 8036:2015(E) 5 Minimum requirements See Table 1. 6 Ingredients Immersion liquids conforming to this International Standard shall not contain ingredients which are known to have any adverse eff
33、ects on safety or health. 7 Designation of immersion liquids Immersion liquids conforming to this International Standard shall be designated as in the following example: EXAMPLE Immersion liquid ISO 8036 Type F. 8 Labelling of containers Immersion liquids are commonly packed and marketed in brown gl
34、ass bottles or in oilers made from polyethylene or polypropylene. The labelling of these containers shall include the following information: the product name; the designation in accordance with this International Standard; labelling required by regulations for hazardous freight; the refractive index
35、, n e , at 23 C; the Abbe number, v e ; the batch number; the expiration date when stored in the unopened original 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
36、country of origin if different from that given in the manufacturers address. Table 1 Minimum requirements for immersion liquids a Parameter Type N (standard) Type F ( f o r f l uo r e s c e n c e ) Type G (glycerol) b Reference temperature (C) Tolerance (C) 23 0,1 23 0,1 23 0,1 Spectral range (nm) 4
37、00 to 900 320 to 1 100 230 to 1 100 Refractive index, n e 1,518 0 0,000 5 1,518 0 0,000 5 1,450 0,005 Abbe number, v e 43 4 43 4 58 4 Transmittance (d = 10 mm) at 1 100 nm 80 % 70 % aImmersion liquids, with the exception of glycerol, may not contain volatile or hygroscopic ingredients.Specifications
38、 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 depend 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 I
39、SO 8036:2015ISO 8036:2015(E) Parameter Type N (standard) Type F ( f o r f l uo r e s c e n c e ) Type G (glycerol) b at 1 000 nm 80 % 70 % at 900 nm 90 % 90 % 90 % at 800 nm 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 7
40、5 % 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 % Aut o f l u o r e s c en c e (mg/l quinine sulfate) F(365 nm/450 nm) c 0,06 0,02 F(405 nm/485 nm) c 1,20 0,50 F(313 nm/45
41、0 nm) 0,05 Viscosity (mm 2 /s) at 23 C 0,1 C 50 to 1 500 50 to 1 500 b 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 hygr
42、oscopic 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 sheet The manufacturer or the supplier of an immersion liquid shall make a technical data sheet available on request. This data
43、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 measurements; spectral range;Table 1 (continued) 4 ISO 2015 All rights reservedBS IS
44、O 8036:2015ISO 8036:2015(E) refractive indices n e , n D(at sodium D-line), n g(at mercury g-line), n F , n C ; Abbe number, v e ; temperature coefficient of refractive index in the range 15 C to 40 C; transmittance within the spectral range for an optical path length d = 10 mm; autofluorescence; be
45、haviour 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 disposal. ISO 2015 All rights reserved 5BS ISO 8036:2015ISO 8036:2015(E) ISO 2
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