Showing papers in "Journal of Histochemistry and Cytochemistry in 1966"
TL;DR: The early stages of absorption of intravenously injected horseradish peroxidase in proximal tubules of mouse kidney were studied with a new ultrastructural cytochemical technique, which gives sharp localization and is sensitive to protein transport.
Abstract: The early stages of absorption of intravenously injected horseradish peroxidase in proximal tubules of mouse kidney were studied with a new ultrastructural cytochemical technique. In animals killed as early as 90 sec after injection, reaction product was found on the brushborder membranes and in the apical tubular invaginations. From the latter structures it was transported to the apical vacuoles, in which it was progressively concentrated to form protein absorption droplets. The method, which employs 3,3'-diaminobenzidine as oxidizable substrate, gives sharp localization and is sensitive. This system is advantageous in studying the early stages of renal tubular protein absorption, since small amounts of protein on membranes and in tubules and vesicles can be detected easily. The method also appears promising for studying protein transport in a variety of other cells and tissues.
6,495 citations
TL;DR: The peroxidase was conjugated to antibody by a method similar to one developed for the comijugations of ferritimi to auitibody, and the best amid most consistent resumlts were obtained when FNPS was umsed.
Abstract: primiciples of immummiohiistochemist ry to electron niicroscopy have niet with very limited sumccess (Pierce et at., mt. Rev. Exp. Path. 3: 1. 1964). Ins ami effort to obviate somuie of these himnitatiomus, tissumes have beemt staimied with emszymes conujumgated to antibody. Ins the imuitil experintemits, acid phosphsatase was comijugated to antibody (Ram et al., Fed. Proc. 25: 732. 1966). Although these preliminary experiments deniomustrated the feasibility of thie approachu, the resumlts of the couijugatiomss were so variable that search was made for a better enzyme. Imu the presemut stumdy horseradish peroxidase was employed becaumse the enzynte is commercially available ims relatively pure form; the cytochemical amid histochemical methods have been well established, and successfully adapted to electron microscopy (Graham amid Karnovsky, J. Histochem. (Jytochem. 14: 291. 1966). The peroxidase was conjugated to antibody by a method similar to one developed for the comijugations of ferritimi to auitibody (Ram et at., J. Cell Biol. 17: 673. 1963), amid acid phosphiatase to amitibody (Ram et at., Fed. Proc. 25: 732. 1966; Nakane et at., J. Histochein. Cytochem. 7th Anmuumal Meetimig, Abstract, 1966), which employed the bifummictional reagent, p,p’difiumoro-m-m’-diuiitrodiphenyl sulfomie (FNPS). Other bifummuctiomual reagemuts sucht as 1-ethiyl-3-(3dimethylanuino propyl) carbodiimide have also beemi employed, bunt the best amid most consistent resumlts were obtained when FNPS was umsed. For routimie stumdies the conjumgates were pre-
1,005 citations
TL;DR: In discussing the morphological alterations of the liver induced by phenobarbital, numerous biochemical changes reported in the literature have been reviewed in an effort to gain a clearer view of the over-all significance of the agranular reticulum in liver cell function.
Abstract: The electron microscopic appearance of normal hamster liver is illustrated, and attention is drawn to certain unusual features of hepatocyte fine structure that have not hitherto been described. Ex...
291 citations
TL;DR: Localization of two soluble compounds,3H-estradiol in liver and uterus and 3H-mesobilirubinogen in liver, has been studied by six autoradiographic methods with varying degrees of diffusion.
Abstract: Localization of two soluble compounds, 3H-estradiol in liver and uterus and 3H-mesobilirubinogen in liver, has been studied by six autoradiographic methods: (I) Frozen sections, freeze-dried, dry mounted on emulsion-coated slides; (II) Frozen sections thawed on emulsion-coated slides; (III) Frozen sections thawed on glass slides and dipped in liquid emulsion; (IV) Freeze-dried tissue, vapor-fixed, epoxy-embedded, sectioned, dipped in liquid emulsion; (V) Freeze-dried tissue, paraffin-embedded, sectioned, deparaffinized, dipped in liquid emulsion; and (VI) Formalin-fixed tissue, paraffin-embedded, sectioned, deparaffinized and dipped in liquid emulsion. Methods II-VI show varying degrees of diffusion, and in some cases loss of activity from tissue, when compared with results obtained by dry mounting of freeze-dried frozen sections. Autoradiograms prepared by dry mounting of 0.75-, and 1-µ thick sections, cut and dried below –60°C, have been found superior.
259 citations
TL;DR: Lungs from marsupials, bats and rodents were studied by light and electron microscopy and it was proposed that these bodies are the precursors of cytosomes.
Abstract: Lungs front marsupials, bats and rodents were studied by light and electron microscopy. In all three groups, the great alveolar cells exhibit similar morphologic and cyt.ochemical characteristics. Cytoplasmic vacuoles seen its these cells by light microscopy correspond to cytosomes that are demonstrable in them by electron microscopy. Such cytosomes are osniiophilic, periodic acid-Schiff -positive amid stainable with Sudan black after acetone extraction. After fixation in a mixture of aldehydes, followed by extraction its chloroform-methanol and postflxation in osmiuni let roxide, cytosonies lose their osmiophilia. The cytoplasm of the great alveolar cell is notable for a loosely ordered granular endoplasmic reticulum, an extensive Golgi apparat.us and numerous iii ivesicular bodies. Many forms transitional in appearance between mult ivesicular bodies and cytosomes are present. In these, osmiophilic matter occupies the intervesicular space. It. is proposed that these bodies are the precursors of cytosomes. The cytosomes are imit.erpreted to be products of the “lysosomal” system in this cell. Ultimately they are secreted onto the alveolar surface.
256 citations
TL;DR: A previously undescribed catalytic action of lead ion on the nonenzymatic hydrolysis of nucleoside phosphates has been demonstrated and it is suggested that this reaction may be a source of artifact in the lead salt method for the histochemical localization of nucleOSide phosphatases.
Abstract: A previously undescribed catalytic action of lead ion on the nonenzymatic hydrolysis of nucleoside phosphates has been demonstrated. Lead ion (3.6 mM) hydrolyzed adenosine triphosphate (ATP) at pH ...
161 citations
TL;DR: Two methods of Feulgen hydrolysis were examined and it was suggested that depurination is dependent primarily on acid rather than heat, while further degradation is based on heat rather than acid.
Abstract: Two methods of Feulgen hydrolysis (5.0 N HCl at room temperature and 1.0. N HCl at 60°C) were examined by serial microspectrophotometric measurements of the relative DNA content of normal human lym...
157 citations
TL;DR: In Gomori ‘5 met hod (Microscopic Histochernisiry, Ummiversity of Chicago Press, 1958) a precipitate of calcinsm phosphate is produced at pH 9.4 which is thems transformed imito cobalt phosphate which appears as umeedle-like particles.
Abstract: 1)ifferenst techumics have beenm employed for the electron microscopic visualizationi of alkaliume phosphatase activity. These are based oti direct or iumdirect precipitationi of heavy metals. In Gomori ‘5 met hod (Microscopic Histochernisiry, Ummiversity of Chicago Press, 1958) a precipitate of calcinsm phosphate is produced at pH 9.4 which is thems transformed imito cobalt phosphate. The deposits appear as umeedle-like particles. The localization is niot too precise, owing to the incomplete tranmsformatioum of calcium phosphate itmto cobalt phosphate. Iii a slight modification of this method, de Th (6#{232}ineColl. .4nn. Soc. Fr. Microsc. Electron., Marseille, 40, 1965) recently used lead isitrate iusstead of cobalt miitrate amid obtaimmed better results. Ins 1960 M#{246}lbert et al., (J. Biophys. Biochern. (Jytol. 7: 387) described a direct niethod usiumg lead as captnnriumg reagetit. Its order to keel) sufficient amouumts of lead mm solution ium the Tris biuffer, a chelatiumg agent anmd disodimnm pheumyl-
154 citations
TL;DR: Evidence is presented that monocytes, as well as mature neutrophils and their precursors extending back to the progranulocyte, contain significant amounts of this enzyme, and a rare mature eosinophil demonstrated a trace of lysozyme activity.
Abstract: By means of an indirect histochemical technique, the intracellular lysozyme of the formed elements of the peripheral blood and bone marrow was estimated. Evidence is presented that monocytes, as well as mature neutrophils and their precursors extending back to the progranulocyte, contain significant amounts of this enzyme. A rare mature eosinophil demonstrated a trace of lysozyme activity. There was no evidence of lysozyme activity in basophils, erythrocytes, megakaryocytes, platelets, plasma cells, tissue mast cells or bone marrow reticuloendothelial cells.
153 citations
TL;DR: A direct lead method for the visualization of alkaline phosphatase activity at pH 9 was used in a study on the absorptive cells of the duodenum of the mouse and lead phosphate precipitate was observed in several structures after a short incubation of glutaraldehyde-fixed frozen sections.
Abstract: A direct lead method for the visualization of alkaline phosphatase activity at pH 9 was used in a study on the absorptive cells of the duodenum of the mouse. Lead phosphate precipitate was observed in several structures after a short incubation of glutaraldehyde-fixed frozen sections. On microvilli and in vacuoles below the terminal web, the reaction was intense and persisted even after osmic acid fixation. In the Golgi zone, in the smooth reticulum cisternae and in the dense bodies, the reaction was prominent. In these sites, the deposits were not observed after osmic acid fixation or after short incubation in media containing substrates other than β-glycerophosphate or α-naphthylphosphate.
152 citations
TL;DR: The amount of lead-catalyzed hydrolysis was in the same order of magnitude as fixed tissue ATPase activity and could quantitatively account for the amount of phosphate needed to give recognizable localization of lead salt deposits in sections of fixed tissue.
Abstract: The lead method of Wachstein and Meisel for the histochemical localization of adenosine triphosphatase (ATPase) involves the incubation of sections of fixed tissue in reaction mixtures containing ATP, lead nitrate, magnesium sulfate and a Tris-maleate buffer, pH 7.2. Both fixation and the presence of lead ion were shown to inhibit tissue ATPase activity markedly and to inactivate the sodium- plus potassium-dependent membrane ATPase. In addition, recent studies have demonstrated that lead ion, in the concentration used in the Wachstein-Meisel system, will catalyze the hydrolysis of ATP. Studies on the effect of this nonenzymatic reaction on the histochemical localization of ATPases demonstrated that plasma membrane localization occurred only with lead and ATP concentrations which gave significant nonenzymatic hydrolysis of ATP by lead. In addition, nuclear and mitochondrial localization without accompanying plasma membrane localization could be obtained in formalin-fixed tissue with decreased concentration...
TL;DR: The characterization of human lipofuscin pigment granules as lysosomes is strengthened by the cytochemical demonstration of β-glucuronidase and glucosaminidase activities in hepatocyte pigment granule which also have acid phosphatase activity as mentioned in this paper.
Abstract: The characterization of human lipofuscin pigment granules as lysosomes is strengthened by the cytochemical demonstration of β-glucuronidase and glucosaminidase activities in hepatocyte pigment granules which also have acid phosphatase activity. Lysosomes of parenchymal cells and macrophages are seen to differ in their content of cytochemically demonstrable hydrolase activities. Reduced diphosphopyridine nucleotide tetrazolium reductase and peroxidase activities that are remarkably resistant to heat (90°C for 15 min) and prolonged fixation are also visualized in lipofuscin granules. It is suggested that these oxidative activities reflect the presence of heme compounds that accumulate during the degradation of cytoplasmic constituents in autophagic vacuoles or dense bodies. The presence of metabolites with peroxidase activity within lysosomes may be responsible for the oxidation of melanin precursors to melanin and unsaturated fat to the alcohol-insoluble lipid of lipofuscin pigment. Cytoplasmic iron (hemos...
TL;DR: By means of a mechanical coating instrument a fast, simple method to coat specimens with liquid nuclear track emulsion has been devised for quantitative light and electron microscopic radioautography.
Abstract: By means of a mechanical coating instrument a fast, simple method to coat specimens with liquid nuclear track emulsion has been devised for quantitative light and electron microscopic radioautography. In both cases, the section is mounted on a glass slide. After the vertically held slide has been immersed in the melted emulsion, the instrument withdraws it at a slow, constant speed. As a result, the specimen is coated with a thin, uniform emulsion layer composed of homogeneously distributed silver bromide crystals. The thickness of the emulsion coat may be standardized by selection of an optimal combination of emulsion dilution, temperature and withdrawal speed.
TL;DR: Protection of the enzyme activity was afforded by other polyphosphates, particularly sodium pyrophosphate and, to less degree, thiamine pyroph phosphate, adenosine diph phosphate and uridine triphosphate.
Abstract: Studies have been made on the effect of fixatives containing substrate and other additives on the histochemical staining reaction for myosin adenosine triphosphatase activity in the leg muscle, and α-glycerophosphate and succinate dehydrogenase activities in the kidney of the rat. Myosin adenosine triphosphatase was well preserved in the presence of adenosine triphosphate in fixative prepared from methanol-free formaldehyde and buffered to pH 7.2 with cacodylate. Addition of sucrose was found to increase the enzyme preservation. Similar protection of the enzyme activity was afforded by other polyphosphates, particularly sodium pyrophosphate and, to less degree, thiamine pyrophosphate, adenosine diphosphate and uridine triphosphate. α-Glycerophosphate dehydrogenase was also preserved to greater degree by a similar fixative containing α-glycerophosphate than by substrate-free fixative. Succinate dehydrogenase was not significantly preserved in succinate-containing fixative. On the other hand, sucrose in the...
TL;DR: Study of fresh blood and tissues from man and many animals revealed histamine its the mast cells, blood vessels, gastric mucosa, and in some species, platelets.
Abstract: A simple, rapid technique has been developed for the visualization of histamine in its cellular depots throughout the body. Based on the yellow fluorochrome formed by the interaction of o-phthalald...
TL;DR: The soleus, sartorius and brachioradialis from 10 other species of primates had the same relative succinic dehydrogenase activities and histochemical staining patterns as the rhesus.
Abstract: There was a direct correlation between the qualitative histochemical classification by staining intensity for succinic dehydrogenase and the quantitative measurements of succinic dehydrogenase activity for the quadratus femoris (red), soleus (red), sartorius (predominantly red) and the superficial portion of the brachioradialis (predominantly white) muscles of the rhesus monkey. The relative succinic dehydrogenase activities were quadratus femoris > soleus > sartorius > brachioradialis, the quadratus femoris having 7 times more enzyme activity than the brachioradialis. The sartorius of male rhesus monkeys had a higher enzyme activity than that of the female. Muscle samples were stained with sirius red and graded for amounts of connective tissue as follows: soleus < sartorius < brachioradialis. These histochemical results were verified by chemical analyses. The soleus, sartorius and brachioradialis from 10 other species of primates had the same relative succinic dehydrogenase activities and histochemical s...
TL;DR: The inability to demonstrate the hepatic enzyme histochemically is believed to depend on the lower concentration of the enzyme in liver as compared with its concentration in the duodenal mucosa.
Abstract: A comparative study of xanthine oxidase in rat duodenum and liver was carried out by a histochemical techniquie, by cell fractionation and zone electrophoresis. With the histochemical method presented, reaction produuct could be demonstrated in the epithehial cells of the duodenal vihli in glintanaldehyde-fixed cryostat sections incubated in 0.2 M sodium phosphate bunffer at pIT 7.4 containing 2.5 X 10� M hypoxanthine and 1 mg/mi nitro-bluie tetrazolium (Nitro-BT). No reaction prodtnct counld be observed, however, in similarly treated sections of rat liven. Liver and dunodenal homogenates showed high enzyme activity when assayed by a microfiunorometric method using 2-amino-4-hydroxy pteridine as substrate. Fractionation of these homogenates revealed the presence of enzyme activity only in the high speed supernatant fraction. This fraction was also studied by zone electrophoresis on vertical starch gel and by acrylamide gel, disc ehectnophoresis. Xanthine oxidase separated electrophoreticaily into two distimict bands and was localized by incunbation of the gels in the same medium as described for t.he histochemical method. The duodenal enzyme showed a more rapid mobility than that of the liver. The imiabihity to demonstrate the hepatic enzyme histochemically is believed to depend on the lower concemitration of the enzyme in liver as compared with its concentration in the duodenal mucosa.
TL;DR: Post-formalin ammoniacal-silver, a staining method which is selective for histone, imparts different colors, ranging from yellowish to black, to many different cell types, and the study of histone specificity facilitated.
Abstract: Post-formalin ammoniacal-silver, a staining method which is selective for histone, imparts different colors, ranging from yellowish to black, to many different cell types. The same colors are imparted to histone extracts of the cell types in question. Pure lysine as well as so-called lysine-rich histone subfractions stain yellowish, while pure arginine and so-called arginine-rich subfractions stain black. The mechanism of the color differences of the stain is somewhat elucidated by these findings, and the study of histone specificity facilitated.
TL;DR: It was found by optical microscopy that rabbit pancreatic islet cell acid phosphatase activity is present in discrete, mostly perinuclear foci and that this distribution differs from that of the aldehyde fuchsin-positive secretory granules which are densely packed at the capillary pole of the cell.
Abstract: Utilizing formaldehyde- or glutaraldehyde-fixed tissue and Gomori's lead method it was found by optical microscopy that rabbit pancreatic islet cell acid phosphatase activity is present in discrete...
TL;DR: Stochastic localization of cholinesterase activity was demonstrated in rat sciatic nerve using thiocholine esters as substrate and studies of specific inhibition sensitivities and substrate preferences suggested that the axonal choliersterase of rat Sciatic nerve is a specific acetylcholinestersterase which is relatively homogeneous.
Abstract: Ultrastructural localization of cholinesterase activity was demonstrated in rat sciatic nerve using thiocholine esters as substrate. This enzymatic activity was limited to axons, where it was located on limiting axonal membranes, and on axonal vesicles of all unmyelinated and some myelinated nerve fibers. No accentuation of activity was present at nodes of Ranvier. The use of acetyl, propionyl and butyrylthiocholine as substrate resulted in decreasing amounts of end product precipitate, respectively. The cholinesterase activity was sensitive to heat, resistant to N-ethyl maleimide, and inhibited byeserine, DFP, E600, RO2-1250 and BW 62C47. Studies of specific inhibition sensitivities and substrate preferences suggested that the axonal cholinesterase of rat sciatic nerve is a specific acetylcholinesterase which is relatively homogeneous.
TL;DR: The significance of the presence and localization of catecholamines in the toad bladder is discussed in relation to the pharmacology of the organ and to histochemical investigations of the innervation in other vertebrate bladders.
TL;DR: Combined electron microscopic and quantitative chemical analysis of the subcellular fractions suggested that some bound nonspecific esterase may be localized in sub-synaptic membranes, which may be determined in part by the participation of acetylcholinesterase in hydrolysis of the substrate.
Abstract: The localization and properties of soluble and bound esterases of subcellular fractions of rat brain have been investigated. Bound esterases were extracted with 1% Triton X-100 and separated by starch gel electrophoresis. By these means a molecular population of isoenzymes was demonstrable that was quantitatively different from the isoenzyme population of the watersoluble esterase activity. The highest specific activity for α-naphthyl acetate hydrolysis was contained in the microsomal fraction and could be extracted by Triton. In contradistinction to whole brain and to other subcellular fractions, microsomes contained a molecular population of esterase isozymes which was qualitatively distinct from that of water extracts in that the fast moving A-type esterases were absent. In addition, there was present a heavy concentration of slow movinig B-esterase. Acetylcholinesterase could also be extracted from this fraction by Triton, migrated with B-esterase and actively hydrolyzed αnaphthyl acetate. Combined el...
TL;DR: It was concluded that in the system investigated the "lyo" and "desmo" components represented, in reality, distinct enzymes with unique characteristics.
Abstract: In order to determine the relationship and characteristics of the soluble and tissue-bound (so-called "lyo" and "desmo") components of enzymes in histochemical and biochemical systems, a biochemical investigation of the hydrolysis of l-leucyl β-naphthylamide in rat kidney was undertaken. By ultracentrifugation, autolysis, salt and solvent fractionation procedures and DEAE-cellulose column chromatography primary soluble and tissuue-bound enzymes were separated and partially purified. The soluble enzyme was found to be inhibited by p-chloromercuribenzoate and reactivated by cysteine. The tissue-bound enzyme was found to be inhibited by o-phenanthroline and reactivated by Co++, and hydrolyzed peptides and amino acid β-naphthylamides at rates differing markedly from those of the soluble enzyme and commercial leucine aminopeptidase. Disc electrophoresis of the tissue-bound enzyme preparation demonstrated two protein bands, both revealing enzymic activity with almost identical hydrolytic characteristics, i.e., ...
TL;DR: High resolution autoradiography of tritium-labeled glucose incorporation offered some further illustration on the process of glycogen formation, including a probable role of this organelle in glycogen synthesis and breakdown.
Abstract: The relation between agranular reticulum and glycogen was studied in hepatic cells of female rats. Prolonged fasting of nonadrenalectomized rats did hot result ins complete liver glycogen depletion, whereas in adrenalectomized rats this could be accomplished within a few hours of fasting. In nonadrenalectomized rats a marked development of agranular reticulum associated with glycogen was found, whereas in adrenalectomized rats no such marked development of agranular reticulum was seen during glycogen depletion. Early glycogen restoration in glycogen-depleted liver cells of adrenalectomized rats was brought about 2-4 hr after the injection of 1 dose of cortisone or ½-1 hr after the injection of glucose. Early restoration of glycogen was accompanied and even preceded by a marked development of tubular agranular reticulum. A probable role of this organelle in glycogen synthesis and breakdown is discussed. High resolution autoradiography of tritium-labeled glucose incorporation offered some further illustrati...
TL;DR: Early concepts of amyloid are reviewed in context with chemical literature from 1839-1859 and it appears possible that investigations of the carbohydrates at sites ofAmyloid formation would aid the understanding of the pathogenesis of ameloidosis.
Abstract: Early concepts of amyloid are reviewed in context with chemical literature from 1839-1859. Cellulose was first described in 1839. Treatment with sulfuric acid converted cellulose into a substance w...
TL;DR: It is concluded that the radioactivity which occurs in osteoblasts, odontoblasts and some fibroblasts soon after injection of 3H-glycine and3H-proline is composed in about equal parts of collagen and of other protein(s).
Abstract: Sections of Canmnoy-fixed tissunes from young rats and adult mice were inctubated for 5 hr at 37#{176} C in a 0.1% soluntiomi of Clostridium histolyticum collagenase punrified by ion exchauuge chromatography. This treatment caunses collagenous fibers as well as dentinal and bone matrices to lose their ability to stain-n red with the van (ieson technique and pink with eosin. In view of the high specificity of the em-nzyme, it is assumed that the stained component of these tissutes is collagen. Immediately after injection of H-glycine or 3H-prohine, radioautography shows the label in the cytoplasm of several cell types: osteoblasts of femur and alveolar bone, odontobiasts of growing molar am-ndiu-ncisor teeth. and fibroblasts of the periodontal membrane. Later, the label appears outside the cells, that is, in bone matrix, dentinal matrix and intercellular spaces of periodontal membram-ne, respectively. Collagenase treatment extracts about half of the label which is present in the cells soon after injection, all or nearly all the label which appears later in bone and dentinal matrix, bunt only a fraction of that which appears in the intercellutlar spaces of the periodontal niennbnane. It is conclunded that the radioactivity which occturs im-n osteoblasts, odonitoblasts and son-ne fibroblasts soon after injection of 3H-glycine and 3H-proline is composed in about equal pants of collagen and of other protein(s). In bone and dentinal matrix collagen alone seems to be deposited, whereas in-nthe periodontal membrane collagen is laid down in association-n with other protein(s).
TL;DR: A histochemical method for triglyceride esters is described that depends on hydrolysis of triglycerides to fatty acids by pancreatic lipase, precipitation of the released fatty acid as calcium soap and formation of lead sulphide by the conventional Gomori technique.
Abstract: A histochemical method for triglyceride esters is described that depeusds on hydrolysis of triglycerides to fatty acids by pancreatic lipase, precipitation of the released fatty acid as calcium soap and formation of lead sulphide by the conventional Gomori technique. The specificity of the lipase for triglyceride was investigated by chromatography arid activatiout-inhihition studies. The eutzyme preparation used hydrolyzed triglycerides and waxes but did utot split cholesterol esters or phosphoglycerides. Positive histochemical reactions were obtaiumed with this pancreatic lipase method in medium sized and fine lipid droplets in frozen sections. Positive reactions were observed in rat brown fat; in various adipose tissues in maui, rabbit and rat; in the spermatogonia of rat testis; in human epidermis and sebaceotts glands; its solitary uuiidentified adrenocortical cells in man attd rat; in human fatty liver and in flume droplets in semtile human myocardium. A slight variable reaction was obtained in humati atherosclerotic plaques. The reaction of brown fat. was also investigated with the electron microscope. No triglyceride was demonstrated in unfixed tissue sectons when an active preparation of lipoprotein lipase was used in place of pancreatic lipase. This failure was probably due to diffttsion of lipoprotein from unfixed sections. Lipoprotein lipase is itiactive agaimmst serum lipoprot.eins fixed with fornialdehyde or glutaraldehyde. Until now the specific histochemical demonstration of triglyceride esters has not been possible. Although it iswidely held that the red Sudan dyes (including oil red 0) are specific
TL;DR: In both porcine and bovine pancreas, the acinar cells were virtually all similar, indicating functional similarity to the extent of these observations.
Abstract: The localization of α-amylase in the pig pancreas, and of chymotrypsinogen and trypsinogen in the beef pancreas, has been investigated by immunofluorescence, using antisera prepared in rabbits against commercial crystalline enzyme preparations. Porcine α-amylase was abundant in almost every acinar cell, being distributed as fine granular material in the basal part of the cell, and also in some of the zymogen granules in the juxta-nuclear region. It was visible also in the Golgi zone. Bovine chymotrypsinogen and trypsinogen were present in the presumably less mature zymogen granules in the cytoplasm around the nucleus, but the granules near the lumen failed to react with the antisera, perhaps because they were hidden by superficial material. The specificity of the anti-amylase serum was high, that of the anti-chymotrypsinogen and anti-trypsinogen lower because of cross reactions with each other. In both porcine and bovine pancreas, the acinar cells were virtually all similar, indicating functional similari...
TL;DR: The resolving power, sensitivity and latent image fading have been studied in autoradiographs prepared by a technique that retains soluble compounds and it was revealed that –25°C was the optimum exposure temperature.
Abstract: The resolving power, sensitivity and latent image fading have been studied in autoradiographs prepared by a technique that retains soluble compounds. A resolution of 2-4 µ was obtained in preparations containing 3H-thymidine and 9-13 µ in those containing 22NaCl. Results showed that there was no significant diffusion of soluble compounds. Sensitivity studies revealed that –25°C was the optimum exposure temperature. There was no significant latent image fading in preparations exposed for periods up to 330 days.