ISSN:0368-3974    

DOI:10.1111/j.1365-2818.1967.tb04500.x

出版商:Blackwell Publishing Ltd    

年代:1967

数据来源: WILEY

摘要:

SYNOPSISThe usual manual compensator of a double‐refracting interference microscope is replaced by a modified version which is continuously driven by a synchronous electric motor. The resulting phase excursions cover a range of a complete light wave‐length and are linearly related to time, so that the light intensity discharged by the microscope fluctuates sinusoidally. When an optical‐phase object in the interference microscope system displaces the phase relationship between the two interfering beams, the resulting phase displacement of the sinusoidally fluctuating light intensity causes the sinusoidal voltage from a photodetector to suffer an equal and corresponding phase displacement. This last displacement is transformed into a directly phase‐proportionate voltage or pulse duration by means of an electronic phase‐measurin

ISSN:0368-3974    

DOI:10.1111/j.1365-2818.1967.tb04501.x

出版商:Blackwell Publishing Ltd    

年代:1967

数据来源: WILEY

ISSN:0368-3974    

DOI:10.1111/j.1365-2818.1967.tb04502.x

出版商:Blackwell Publishing Ltd    

年代:1967

数据来源: WILEY

摘要:

SYNOPSISThe dry mass of living cell inclusions can be calculated from the concentration of their solids (derived from measurement of their refractive indices), and from their volumes (derived from measurements of their dimensions). The accuracy and advantages of this approach in comparing one type of cell material with another are discussed; some general conclusions about the measurement of cellular constituents with interference microscopy are presented.

ISSN:0368-3974    

DOI:10.1111/j.1365-2818.1967.tb04503.x

出版商:Blackwell Publishing Ltd    

年代:1967

数据来源: WILEY

ISSN:0368-3974    

DOI:10.1111/j.1365-2818.1967.tb04504.x

出版商:Blackwell Publishing Ltd    

年代:1967

数据来源: WILEY

摘要:

SYNOPSISThe use of interference techniques in microscopy provides a useful tool for scientists and engineers in the investigation and measurement of very small changes in height on a reflecting surface. The interference fringes may be regarded as contour lines spaced at half the wave length of the monochromatic light used for illumination. This gives, in effect, a remarkably high magnification in depth and makes possible the assessment of the variations of flatness and finish of highly finished surfaces and the measurement of the thickness of vacuum evaporated films and the height of surface steps on crystals.Replica techniques can be used for the examination of inaccessible surfaces and also for the variation of the value of the fringe spacing if the pellicle is viewed when immersed in liquids of various refractive indices, thus varying the effective magnification and hence the range of measurement.Further, the use of fringes of equal chromatic order, involving the use of a spectroscope as an accessory to the microscope, as described by Tolansky and others, enables depth measurements to an accuracy of a few micrometres to be made readily.

ISSN:0368-3974    

DOI:10.1111/j.1365-2818.1967.tb04505.x

出版商:Blackwell Publishing Ltd    

年代:1967

数据来源: WILEY

摘要:

SYNOPSISMeasurements with the interference microscope show the optical‐path difference relative to the medium of many biological objects after histological processing to be less in water than would be expected on the basis of measurements made in other media. This anomalous optical‐path difference is found also with sections of gelatin (autoradiographic stripping film), but not if the gelatin is mounted on the slide in such a way that swelling is constrained to a direction parallel to the optical axis of the microscope. Swelling in water has been demonstrated in a variety of formalin‐fixed and de‐fatted tissues.It is suggested that the anomalous optical‐path differences in water are due to swelling causing a change in the lateral dimensions of the object, and hence loss of material from the optical path. The fact that the anomaly has also been found with scanning and integrating interference microscopes is discussed, and it is suggested that the validity of such methods needs re‐

ISSN:0368-3974    

DOI:10.1111/j.1365-2818.1967.tb04506.x

出版商:Blackwell Publishing Ltd    

年代:1967

数据来源: WILEY

摘要:

SYNOPSISSulphated mucosubstances in sections of coagulant‐fixed tissue emit a variable yellow to red fluorescence after being stained with purified coriphosphine at pH 2 to 4. Unfortunately this fluorescence is sometimes indistinguishable from a similarly coloured fluorescence emitted by nuclei, oxidized sulphoproteins and some sialo‐mucins, which, however, can be eliminated more or less selectively either by (1) a preliminary Feulgen hydrolysis and condensation with N,N‐dimethyl‐m‐phenylene diamine; or (2) by pretreatment with a solution of ferric alum; or (3) by a previous methylation with methanolic thionyl chloride, followed by reduction with lithium aluminium hydride in hot dioxane and then by saponification (a sequence of reactions which, in theory but not always in practice, should remove all polyanionic groups in the tissueexceptsulphate groups on sulphated mucopolysaccharides); or (4), best of all, by staining sections in a very dilute solution of thiazol yellow at pH 2 after they have been stained in coriphosphine.The rationale and limitations of these methods are discussed c

ISSN:0368-3974    

DOI:10.1111/j.1365-2818.1967.tb04507.x

出版商:Blackwell Publishing Ltd    

年代:1967

数据来源: WILEY

摘要:

SYNOPSISAccording to Kasten, periodate‐oxidized mucosubstances which have been stained with his fluorescent Schiff‐type reagents (solutions of basic fluorescent dyes saturated with sulphur‐dioxide) emit a specific fluorescence in sections of fixed tissue. In my experience it is sometimes difficult to observe this fluorescence because it is obscured by the similarly‐coloured fluorescence emitted by proteins and nuclei. This and other difficulties, both theoretical and practical, can be mostly overcome by methylating tissue sections beforehand with a methanolic solution of thionyl chloride, then oxidizing them in periodic acid, and finally treating them first with sulphurous acid (at pH 3) and second with a solution of any basic fluorescent dye at a pH between 2 and 3. In sections treated in this way, nearly all periodate‐oxidized mucosubstances emit a much more intense fluorescence than when treated with Kasten's Schiff‐type reagents; and furthermore, that such mucosubstances fluoresce at all lends support to the theory that aldehydes react with Schiff's reagent proper not through N‐sulphinic acids, as has been commonly supposed, but by reacting first with the sulphurous acid present in Schiff's reagent to form an intermediate alkyl sulphonic acid which then combines with basic pararosaniline (the essential ingredient of Schiff's reagent) to give the familiar magenta‐c

ISSN:0368-3974    

DOI:10.1111/j.1365-2818.1967.tb04508.x

出版商:Blackwell Publishing Ltd    

年代:1967

数据来源: WILEY

摘要:

SYNOPSISSalicylhydrazide forms blue fluorescent derivatives with all potentially Schiff‐reactive, periodate‐oxidized mucosubstances in sections of fixed tissue. The derivatives formed from sulpho‐ and sialomucins usually emit an intense fluorescence. The fluorescence emitted by neutral mucosubstance derivatives is less intense. Aluminium salts enhance the fluorescence emitted by these derivatives, but zinc salts tend to quench it.Salicylhydrazide forms stable fluorescent products only with aldehydes, and is therefore highly specific. Moreover, its aldehyde hydrazones are unique among aldehyde acid arylhydrazones in that only they can be converted into formazans.Cytoplasmic proteins emit a red or purple fluorescence after being treated with a solution containing a solochrome black dye and an aluminium salt. This fluorescence enables the blue fluorescence emitted by periodate‐oxidized mucosubstance salicylhydrazones to be seen more

ISSN:0368-3974    

DOI:10.1111/j.1365-2818.1967.tb04509.x

出版商:Blackwell Publishing Ltd    

年代:1967

数据来源: WILEY