BS ISO 8036:2015 pdf free.Microscopes一Immersion liquids for light microscopy.
BS ISO 8036 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 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 referenced 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 offlash and fire points — Cleveland open cup method
ISO 8255-1, Microscopes — Cover glasses — Part 1: Dimensionaltolerances, thickness and optical properties
ISO 8255-2, Microscopes — Cover glasses — Part 2: Quality of materials, standards of finish and mode of
packaging
3 Classification
Depending on their field of application, immersion liquids are classified 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.
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 quartz cuvettes with an optical path length through the immersion liquid old = 10 mm against an empty reference cuvette.
4.3 Autofluorescence
The 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 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 emission at 485 nm.
As the fluorescence emission decreases during the measurement, the autofluorescence of immersion liquids is averaged over a measurement time of 60s.
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 the quality standards for fluorescence spectroscopy.
Prepare a quinine sulfate stock solution of 500 mg/I with 0,05 mol/l sulfuric acid. Before use, prepare two standard solutions of 0,1 mg/I and 2,5 mg/I quinine 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 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 the autofluorescence of the immersion liquids is given in terms of the equivalent amount of quinine sulfate, in mg/I.
NOTE 2 Quinine sulfate is more suitable foruse asa 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.BS ISO 8036 pdf free download.
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