Polyacrylamide electrophoresis of peroxidases on microscope slides and evaluation with the aid of reflecting light

The size, shape and surface morphology of human erythrocytes (RBC) were examined by scanning electron microscopy (SEM), of the fixed material directly and by transmission electron microscopy (TEM) of surface replicas to compare the relative merits of these two observational procedures for this type specimen.A sample of human blood was fixed in glutaraldehyde and washed in distilled water by centrifugation. The washed RBC's were spread on freshly cleaved mica and on aluminum coated microscope slides and then air dried at room temperature. The SEM specimens were rotary coated with 150Å of 60:40- gold:palladium alloy in a vacuum evaporator using a new combination spinning and tilting device. The TEM specimens were preshadowed with platinum and then rotary coated with carbon in the same device. After stripping the RBC-Pt-C composite film, the RBC's were dissolved in 2.5N HNO3 followed by 0.2N NaOH leaving the preshadowed surface replicas showing positive topography.

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Electron-Microscopic localization of biotinylated gastrin bound to eosinophils in the rat stomach

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100169237 ◽

1994 ◽

Vol 52 ◽

pp. 300-301

Author(s):

Caroline A. Miller ◽

David H. Nichols ◽

Richard F. Murphy

Keyword(s):

Electron Microscope ◽

Phosphate Buffer ◽

Uranyl Acetate ◽

List Type ◽

Cerium Chloride ◽

Rat Stomach ◽

Electron Microscopic ◽

Human Sequence ◽

Acetate Lead ◽

Microscope Slides

Gastrin is a small peptide capable of both stimulating gastric acid secretion and acting as an enteric growth factor. Known functions of eosinophils in the rat stomach are related to immunological defense. Here we demonstrate the binding of biotinylated gastrin to rat stomach eosinophils in the electron microscope. Small pieces of stomach were fixed by immersion in 4% paraformaldehyde/0.1% glutaraldehyde in 0.1 M phosphate buffer, pH 7.4 for 1 hour. The tissue was then cryoprotected in 30% sucrose/0.1 M phosphate buffer, transferred to Tissue Tek OCT compound and frozen in isopentane cooled with liquid nitrogen. Transverse cryostat sections were cut at 25 μm, thawed in PBS and free floating sections exposed to 10−5 M biotinylated 1-17 gastrin (human sequence; Peninsula Labs) for 1 hour. Controls omitted the biotinylated gastrin from this step. Sections were then rinsed 3X in PBS and exposed to either:1).a 1:50 dilution of 10 nm Extravidin colloidal gold (Sigma) for 2 hours, or2).an avidin-biotin-alkaline phosphatase complex (ABC-AP;Vector) for 1 hour. A substrate solution containing cerium chloride was used to generate an electron dense reaction product.Sections from both procedures were postfixed in 1% OsO4 in 0.1 M phosphate buffer, rinsed and dehydrated. These were then flat embedded in EMbed 812 between two microscope slides coated with Liquid Release (both from Electron Microscopy Sciences).Polymerized sections were adhered to resin blocks using super glue, cut at 70-90 nm, stained with uranyl acetate/lead citrate and observed in a Philips CM-10 electron microscope.

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Automated Microdensitometry And Protein Assays As Measures For Platelet Adhesion And Aggregation On Collagen Coated Slides Under Controlled Flow Conditions

10.1055/s-0038-1665758 ◽

1979 ◽

Author(s):

R Muggli ◽

H Baumgartner ◽

Th Tschopp

Keyword(s):

Surface Coverage ◽

Platelet Surface ◽

En Face ◽

Uncoated Surface ◽

Unit Surface ◽

Perfusion Time ◽

Flowing Blood ◽

Controlled Flow ◽

Protein Assays ◽

Microscope Slides

Microscope slides were homogeneously coated over a length of 2 cm with a mixture of soluble and fibrillar collagen and exposed at 37°C and under laminar flow to citrated whole rabbit blood at a flow-rate of 100 ml/min. Surface coverage with platelets (adhesion) and platelet accumulations higher than about 5 μm in height (aggregation) were determined by automated microdensitometry of fuchsine stained ‘en face’ preparations. The platelet mass per unit surface was measured with a modified Lowry technique whose sensitivity was equivalent to 5×l05platelets. Platelet number, amount of protein and surface coverage with platelet accumulations correlated. After a perfusion time of 10 min thrombi up to 30 μm in height and oriented in the direction of flow had developed on the collagen coated area. Surface coverage with platelets was 75% and the amount of deposited protein 1.4 μg/mm2(2×l06platelets/mm2). On the uncoated surface single platelets predominated; the surface coverage was 20% and the density of platelets 8×104/mm2. Acetyl- salicylic acid at 100 μm decreased platelet aggregation by about 80% without affecting adhesion.The new parallel plate perfusion system offers rapid quantitation of platelet-surface and platelet-platelet interaction after exposure to flowing blood and iftay also be diagnostically useful.

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An apparatus for vertical gel electrophoresis on microscope slides

Clinica Chimica Acta ◽

10.1016/0009-8981(69)90305-2 ◽

1969 ◽

Vol 26 (1) ◽

pp. 174-176 ◽

Cited By ~ 4

Author(s):

A. Chalvardjian

Keyword(s):

Gel Electrophoresis ◽

Microscope Slides

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Polyphenol oxidase activity staining in polyacrylamide electrophoresis gels

Journal of Biochemical and Biophysical Methods ◽

10.1016/s0165-022x(96)01201-8 ◽

1997 ◽

Vol 34 (2) ◽

pp. 155-159 ◽

Cited By ~ 18

Author(s):

Antonio Rescigno ◽

Francesca Sollai ◽

Andrea C Rinaldi ◽

Giulia Soddu ◽

Enrico Sanjust

Keyword(s):

Polyphenol Oxidase ◽

Oxidase Activity ◽

Polyphenol Oxidase Activity ◽

Activity Staining ◽

Polyacrylamide Electrophoresis

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The disposition of benzo(a)pyrene in the periphyton communities of two South Carolina streams: uptake and biotransformation

Canadian Journal of Botany ◽

10.1139/b82-255 ◽

1982 ◽

Vol 60 (10) ◽

pp. 2084-2091 ◽

Cited By ~ 9

Author(s):

Mary G. Bruno ◽

Timothy E. Fannin ◽

Gordon J. Leversee

Keyword(s):

South Carolina ◽

Slide Surface ◽

Uptake Rates ◽

Area Basis ◽

Autoradiographic Analysis ◽

Colonization Time ◽

Microscope Slides ◽

Surface Area Basis ◽

Glass Microscope ◽

Sheath Material

The effect of periphyton community composition and colonization time on the uptake and biotransformation of benzo(a)pyrene (BaP) was determined in laboratory studies. Naturally colonized glass microscope slides were collected after 3 and 6 weeks from Castor Creek, which has a predominantly desmid flora, and after 3 and 5 weeks from diatom-dominated Upper Three Runs Creek. When expressed on a slide surface-area basis, the Castor Creek periphyton showed significantly greater BaP uptake rates at both colonization periods. Within streams, uptake rates increased significantly with colonization time. Autoradiographic analysis suggests that BaP was accumulated by surface sorption, especially to gelatinous sheath material. Active biotransformation as measured by the percentage extractable non-BaP 14C was not detected in either community.

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A Road Section near Guy's Hill, Jamaica

Geological Magazine ◽

10.1017/s0016756800073933 ◽

1942 ◽

Vol 79 (4) ◽

pp. 241-252 ◽

Cited By ~ 3

Author(s):

C. A. Matley ◽

Frank Raw

Keyword(s):

Sedimentary Rocks ◽

Plutonic Rock ◽

Igneous Rocks ◽

The Road ◽

Road Section ◽

Igneous Intrusions ◽

Microscope Slides

The rocks exposed along the road between Linstead and Guy's Hill, Jamaica, were described by Dr. C. T. Trechmann in this magazine in 1936 (pp. 259–260). The chief object of his account was to prove that the igneous rocks there were intrusions later than the associated Cretaceous and Tertiary limestones, which, according to him, had been metamorphosed into hornfelses, some of which, he stated later (1937, p. 561), he knew to have an “igneous” appearance under the microscope, “which tends to support my contention that in Jamaica we have sedimentaries altered in situ into rocks that would ordinarily be classified as igneous.” Dissent from his descriptions and interpretations was expressed by C. A. M. (Matley, 1937, pp. 501–3), the criticisms being mainly based on an examination of Trechmann's own microscope slides by F. R. A visit to Jamaica by C. A. M. in 1939 allowed him to study this road and to collect a suite of rocks for petrological examination. The results show that Trechmann's interpretation cannot be sustained. There is no granodiorite or other plutonic rock present, no metamorphism hornfelsing the sedimentary rocks, and no igneous intrusions into the Tertiary limestones.

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