scholarly journals Statistical approach for subwavelength measurements with a conventional light microscope

1991 ◽  
Vol 60 (5) ◽  
pp. 1147-1155 ◽  
Author(s):  
Daniel Palanker ◽  
Aaron Lewis
Author(s):  
A. Boyde

It has been proposed that an overriding advantage of the “black-box” configuration of confocal scanning laser microscope (CSLM) is that it enables the use of all the familiar and conventional light microscope (LM) modes to discover a location of interest in a prepared, contrived, biological sample and then to switch to the confocal mode when desired. By implication, it has been assumed that this would not be possible in the Tandem Scanning Reflected Light Microscope (TSM). It is shown here that such suppositions are incorrect, and that correlations with different LM modes are very easily achieved in standard TSMs. In fact, because of the slow scanning speed of existing commercial CSLMs, it is not just an advantage to have other modes of image formation available, but a pure necessity, since finding the area of interest would otherwise be too difficult.


Author(s):  
G. Schatten ◽  
S. Paddock ◽  
P. Cooke ◽  
J. Pawley

Confocal microscopy holds great promise for improved imaging of fluorescent or reflective biomedical specimens. The IMR is actively investigating the advantages and optimal usage of the Medical Research Council's Lasersharp laser - scanning confocal microscope and Tracor/Northern's Tandem Scanning Microscope, which benefits from the principles outlined by Petran et al. and Boyde.Quantitative evaluation of microscopic images has always been complicated by the effect of out-of-focus structures on the final image. These effects can be greatly reduced if the conventional light microscope is replaced by a scanning-confocal light microscope. In such an instrument two conditions are met: 1) only a single point of the sample is illuminated at any time and 2) this point on the sample is then imaged onto the pinhole at the entrance to the photodetector. Because little light from out-of-focus planes will pass through the pinhole, only in-focus data is recorded.


2018 ◽  
Author(s):  
A S M Waliullah

Objectives: To check the feasibility of using mobile phone-based microscopy for various types of human histopathological sample investigations. Methodology: A feasibility study was performed by imaging several histopathological samples with one novel type of microscope ‘Foldscope’ and image compared with a conventional microscope in the laboratory facility. The image acquired from both sources were edited further and put together for comparison. Results: Mobile phone-based microscope acquired images were observed and compared with a conventional microscope and found morphology of the tissue sections were significantly similar as of conventional light microscope images. Conclusion: By comparing the image of some non-human histopathological sample, it could be stated that this method is also feasible for human histopathological sample investigations, especially in the low resource area or in case of emergency situations.


2020 ◽  
Vol 43 (3) ◽  
pp. 34-40
Author(s):  
Pakpoom Thintharua ◽  
Permphan Dharmasaroja

Histology is an essential field in the education of medical students, and competent knowledge in histology is very important when studying pathology. Current teaching methods for histology in medical schools involve using a conventional light microscope (CM) with or without a virtual microscope (VM). This review aims to present advantages and disadvantages of using CM and VM in terms of teaching and learning histology in the context of undergraduate medical education. One major advantage of the traditional CM histology learning method in laboratory practice is that this allows students to practice using a light microscope; however, study flexibility is limited as the students cannot take the microscope back home for self-study after the histology class has finished. Costly repairs and maintenance must also be considered when using CM. By contrast, VM technology can provide flexibility and convenience for both students and staffs. This method allows students to both self-study and group-study almost anywhere at any time. This review emphasizes that histology learning in undergraduate medical education using VM is no longer confined to the classroom. However, the basic skill of how to operate a conventional light microscope is still important for medical students because CM is commonly used in the hospital laboratories and some hospitals may not be equipped with VM technology.  


Author(s):  
W. H. Abbott ◽  
C. O. Pollard

Frustules of the freshwater diatom Melosira granulata (Ehr.) Ralfs from a freshwater diatomite of Miocene age and recent frustules of this species from Reelfoot Lake, Tennessee were examined under a scanning electron microscope to determine fine structure generally not seen or poorly seen with the conventional light microscope.The terminology used to describe frustule structure in this study was originated by Hendy.Melosira granulata generally grows in chains of two connected frustules with each frustule containing two valves or halves. The connection between frustules is accomplished by the encasing of the adjacent valves of two frustules by a structure called the girdle (Fig. 1a).


1967 ◽  
Vol 2 (2) ◽  
pp. 163-168
Author(s):  
J. G. GALL

The analysis of electron micrographs by optical diffraction was introduced recently by Klug & Berger (1964). Their experiments were conducted with a special diffractometer designed for use with diffracting masks up to several inches in diameter. A method is described here for using a conventional light microscope as an optical diffractometer which can accept masks up to 5 mm in diameter. A 100-µ electron-microscope aperture is used as a pinhole source of illumination, and the micrograph to be studied is introduced above the objective. The diffraction pattern produced by the micrograph appears in the usual image plane of the microscope within the eyepiece.


Author(s):  
B. J. Panessa ◽  
J. F. Gennaro

Tissue from the hood and sarcophagus regions were fixed in 6% glutaraldehyde in 1 M.cacodylate buffer and washed in buffer. Tissue for SEM was partially dried, attached to aluminium targets with silver conducting paint, carbon-gold coated(100-500Å), and examined in a Kent Cambridge Stereoscan S4. Tissue for the light microscope was post fixed in 1% aqueous OsO4, dehydrated in acetone (4°C), embedded in Epon 812 and sectioned at ½u on a Sorvall MT 2 ultramicrotome. Cross and longitudinal sections were cut and stained with PAS, 0.5% toluidine blue and 1% azure II-methylene blue. Measurements were made from both SEM and Light micrographs.The tissue had two structurally distinct surfaces, an outer surface with small (225-500 µ) pubescent hairs (12/mm2), numerous stoma (77/mm2), and nectar glands(8/mm2); and an inner surface with large (784-1000 µ)stiff hairs(4/mm2), fewer stoma (46/mm2) and larger, more complex glands(16/mm2), presumably of a digestive nature.


Author(s):  
J. G. Adams ◽  
M. M. Campbell ◽  
H. Thomas ◽  
J. J. Ghldonl

Since the introduction of epoxy resins as embedding material for electron microscopy, the list of new formulations and variations of widely accepted mixtures has grown rapidly. Described here is a resin system utilizing Maraglas 655, Dow D.E.R. 732, DDSA, and BDMA, which is a variation of the mixtures of Lockwood and Erlandson. In the development of the mixture, the Maraglas and the Dow resins were tested in 3 different volumetric proportions, 6:4, 7:3, and 8:2. Cutting qualities and characteristics of stability in the electron beam and image contrast were evaluated for these epoxy mixtures with anhydride (DDSA) to epoxy ratios of 0.4, 0.55, and 0.7. Each mixture was polymerized overnight at 60°C with 2% and 3% BDMA.Although the differences among the test resins were slight in terms of cutting ease, general tissue preservation, and stability in the beam, the 7:3 Maraglas to D.E.R. 732 ratio at an anhydride to epoxy ratio of 0.55 polymerized with 3% BDMA proved to be most consistent. The resulting plastic is relatively hard and somewhat brittle which necessitates trimming and facing the block slowly and cautiously to avoid chipping. Sections up to about 2 microns in thickness can be cut and stained with any of several light microscope stains and excellent quality light photomicrographs can be taken of such sections (Fig. 1).


Author(s):  
J. H. Luft

Ruthenium red is one of the few completely inorganic dyes used to stain tissues for light microscopy. This novelty is enhanced by ignorance regarding its staining mechanism. However, its continued usefulness in botany for demonstrating pectic substances attests to selectivity of some sort. Whether understood or not, histochemists continue to be grateful for small favors.Ruthenium red can also be used with the electron microscope. If single cells are exposed to ruthenium red solution, sufficient mass can be bound to produce observable density in the electron microscope. Generally, this effect is not useful with solid tissues because the contrast is wasted on the damaged cells at the block surface, with little dye diffusing more than 25-50 μ into the interior. Although these traces of ruthenium red which penetrate between and around cells are visible in the light microscope, they produce negligible contrast in the electron microscope. However, its presence can be amplified by a reaction with osmium tetroxide, probably catalytically, to be easily visible by EM. Now the density is clearly seen to be extracellular and closely associated with collagen fibers (Fig. 1).


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