A simple method for perfusion fixation of avian liver for electron microscopy

1981 ◽  
Vol 59 (6) ◽  
pp. 1179-1183 ◽  
Author(s):  
M. K. Bhatnagar ◽  
L. L. David ◽  
Olga Vrablic ◽  
A. Therien ◽  
Andre Blouin
Keyword(s):  
Electron Microscopy ◽  
Body Weight ◽  
Flow Rate ◽  
Phosphate Buffer ◽  
Perfusion Pressure ◽  
Maximum Flow ◽  
Compressed Air ◽  
Simple Method ◽  
Dense Matrices ◽  
Perfusion Fixation

A simple method and apparatus are described for perfusion fixation of avian liver for electron microscopy. A constant perfusion pressure is maintained at or below a fixed value with the use of a compressed air cylinder and without the use of automatic devices. A hyperosmotic (580–600 mosm) fixative solution containing 4% glutaraldehyde, 0.0005 M CaCl2, and 0.0005 M MgCl2 in 0.05 M phosphate buffer (pH 7.4, buffer osmolality 122 mosm) produced consistent fixation without swelling or undue shrinkage. The cytoplasmic organelles were well preserved; notably, the mitochondria had electron-dense matrices and well-defined cristae. A pressure of 60 mmHg (1 mmHg = 133.322 Pa) maintained by compressed air permits a minimum to maximum flow rate of 14–19 mL∙min−1∙kg body weight−1, and optimum preservations of the architecture of sinusoids.

scholarly journals Induction of pinocytosis in rat hepatocytes by partial hepatectomy.

1977 ◽  
Vol 72 (3) ◽  
pp. 695-706 ◽  
Author(s):  
M Mori ◽  
A B Novikoff
Keyword(s):  
Electron Microscopy ◽  
Body Weight ◽  
Partial Hepatectomy ◽  
Low Dose ◽  
Periodic Acid ◽  
Light And Electron Microscopy ◽  
Rat Hepatocytes ◽  
Lysosomal Hydrolases ◽  
Periodic Acid Schiff ◽  
Perfusion Fixation

Rat hepatocytes, normally not highly pinocytic cells, becomes so after partial hepatectomy when about two-thirds of the liver is removed. Droplets, up to 20 mum in diameter, develop, initially by addition to smaller pinocytic structures and later by fusion with lysosomes. The droplets contain a material with an electron microscope periodicity characteristic of fibrin; they are periodic acid Schiff-positive as is plasma. It is therefore reasonable to consider plasma glycoproteins to be major components of the droplets. The droplets are at all times membrane delimited, an observation possible only after perfusion fixation. The droplets are positive for three lysosomal hydrolases identified cytochemically: acid phosphatase, N-acetyl-beta-glucosaminidase, and beta-glucuronidase. From light and electron microscopy it is evident that these activities are acquired by fusion with lysosomes, mostly autophagic vacuoles and residual bodies both of which become very numerous after partial hepatectomy. Pinocytic structures are seen relatively infrequently in the hepatocytes of normal rats but a great many are present after partial hepatectomy. They are most easily observed if horseradish peroxidase (HRP) is intravenously injected before sacrifice and sections are incubated for HRP cytochemistry. The low dose of HRP employed (10 mg/100 g body weight) does not induce pinocytosis in controls, either untreated rats or rats subjected to laparotomy, including palpation of the liver. However, in partially hepatectomized rats even a much smaller dose of intravenous HRP (3.3 mg/100 g) visualizes the pinocytic structures in hepatocytes (coated vesicles, channels, cuplike bodies, and droplets). Kupffer cells pinocytose much HRP in both control and partially hepatectomized rats.

Spontaneous Cataract in Nude Athymic Mice

1977 ◽  
Vol 35 ◽  
pp. 512-513
Author(s):  
Ronald H. Bradley ◽  
R. S. Berk ◽  
L. D. Hazlett
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Nude Mouse ◽  
Cellular Immunity ◽  
Phosphate Buffer ◽  
Osmium Tetroxide ◽  
Athymic Mice ◽  
Transmission Microscopy ◽  
Mouse Lens ◽  
Scanning Electron

The nude mouse is a hairless mutant (homozygous for the mutation nude, nu/nu), which is born lacking a thymus and possesses a severe defect in cellular immunity. Spontaneous unilateral cataractous lesions were noted (during ocular examination using a stereomicroscope at 40X) in 14 of a series of 60 animals (20%). This transmission and scanning microscopic study characterizes the morphology of this cataract and contrasts these data with normal nude mouse lens.All animals were sacrificed by an ether overdose. Eyes were enucleated and immersed in a mixed fixative (1% osmium tetroxide and 6% glutaraldehyde in Sorenson's phosphate buffer pH 7.4 at 0-4°C) for 3 hours, dehydrated in graded ethanols and embedded in Epon-Araldite for transmission microscopy. Specimens for scanning electron microscopy were fixed similarly, dehydrated in graded ethanols, then to graded changes of Freon 113 and ethanol to 100% Freon 113 and critically point dried in a Bomar critical point dryer using Freon 13 as the transition fluid.

The Morphology of Vacuolated Cells in the Liver Following Administration of Vitamin A.

1973 ◽  
Vol 31 ◽  
pp. 408-409
Author(s):  
Odell T. Minick ◽  
Hidejiro Yokoo ◽  
Fawzia Batti
Keyword(s):  
Electron Microscopy ◽  
Body Weight ◽  
Vitamin A ◽  
Light And Electron Microscopy ◽  
Liver Cells ◽  
Prominent Feature ◽  
Vascular Space ◽  
Vacuolated Cell ◽  
Vacuolated Cells ◽  
Vacuolated Cytoplasm

Vacuolated cells in the liver of young rats were studied by light and electron microscopy following the administration of vitamin A (200 units per gram of body weight). Their characteristics were compared with similar cells found in untreated animals.In rats given vitamin A, cells with vacuolated cytoplasm were a prominent feature. These cells were found mostly in a perisinusoidal location, although some appeared to be in between liver cells (Fig. 1). Electron microscopy confirmed their location in Disse's space adjacent to the sinusoid and in recesses between liver cells. Some appeared to be bordering the lumen of the sinusoid, but careful observation usually revealed a tenuous endothelial process separating the vacuolated cell from the vascular space. In appropriate sections, fenestrations in the thin endothelial processes were noted (Fig. 2, arrow).

Binucleate Spermiogenesis in the Mouse

1972 ◽  
Vol 30 ◽  
pp. 112-113
Author(s):  
John J. Wolosewick
Keyword(s):  
Electron Microscopy ◽  
Phosphate Buffer ◽  
Seminiferous Epithelium ◽  
Mouse Testis ◽  
Germinal Epithelium ◽  
Intercellular Bridges ◽  
Adult Mice ◽  
Cervical Dislocation ◽  
Human And Mouse

Classically, the male germinal epithelium is depicted as synchronously developing uninucleate spermatids conjoined by intercellular bridges. Recently, binucleate and multinucleate spermatids from human and mouse testis have been reported. The present paper describes certain developmental events in one type of binucleate spermatid in the seminiferous epithelium of the mouse.Testes of adult mice (ABP Jax) were removed from the animals after cervical dislocation and placed into 2.5% glutaraldehyde/Millonig's phosphate buffer (pH 7.2). Testicular capsules were gently split and separated, exposing the tubules. After 15 minutes the tissue was carefully cut into cubes (approx. 1mm), fixed for an additional 45 minutes and processed for electron microscopy.

Turbomolecular Pumped Electron Microscopy of Rubella Virus (In BHK Celis)

1968 ◽  
Vol 26 ◽  
pp. 204-205
Author(s):  
A. B. Taylor ◽  
G. C. Cole ◽  
M. A. Holcomb ◽  
C. A. Baechler
Keyword(s):  
Electron Microscopy ◽  
Rubella Virus ◽  
Phosphate Buffer ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Basal Medium ◽  
Kidney Cells ◽  
Baby Hamster Kidney ◽  
Input Multiplicity ◽  
Baby Hamster Kidney Cells

An aliquot from a continuous fermenter culture of baby hamster kidney cells (BHK-21 Clone PD-4) (Wistar) maintained in Ca free Eagle's Basal Medium containing 2% Kaolin adsorbed fetal calf serum was planted in spinner flasks at 300,000 cells per ml, total volume 600 ml. After equilibration for one day at 35°C to insure that cells were in log phase, the culture was infected with the M-33-AGMK25 BHK-219 strain of rubella at an input multiplicity of about 6 TCID50 per cell. The virus was identified with specific rubella antiserum.Preliminary experiments had shown that such cultures would reach a peak or plateau HA titer of approximately 1:64, 24 hrs after inoculation and would continue to yield virus for 6 to 12 days. One hundred ml aliquot harvests were withdrawn daily and the culture was returned to volume with growth medium and incubation continued. The harvested cells were spun down rapidly at 2500 rpm per 15 mins., fixed in 3.7% gluteraldehyde in Ca free phosphate buffer saline, and post fixed in osmium tetraoxide. After dehydration, the cells were embedded in Epon 812 and cured approximately 20 hrs at 60°C.

Novel epoxy/anhydride alternatives for biological electron microscopy: Physical and performance characteristis of embed 812 and LX 112 in combination with NSA/NMA/DMAE

1989 ◽  
Vol 47 ◽  
pp. 1000-1001
Author(s):  
Joe A. Mascorro ◽  
Gerald S. Kirby
Keyword(s):  
Electron Microscopy ◽  
Flow Rate ◽  
Succinic Anhydride ◽  
Volume Flow Rate ◽  
Mass Density ◽  
Uranyl Acetate ◽  
Volume Flow ◽  
Base Resin ◽  
And Performance ◽  
Acid Anhydrides

Embedding media based upon an epoxy resin of choice and the acid anhydrides dodecenyl succinic anhydride (DDSA), nadic methyl anhydride (NMA), and catalyzed by the tertiary amine 2,4,6-Tri(dimethylaminomethyl) phenol (DMP-30) are widely used in biological electron microscopy. These media possess a viscosity character that can impair tissue infiltration, particularly if original Epon 812 is utilized as the base resin. Other resins that are considerably less viscous than Epon 812 now are available as replacements. Likewise, nonenyl succinic anhydride (NSA) and dimethylaminoethanol (DMAE) are more fluid than their counterparts DDSA and DMP- 30 commonly used in earlier formulations. This work utilizes novel epoxy and anhydride combinations in order to produce embedding media with desirable flow rate and viscosity parameters that, in turn, would allow the medium to optimally infiltrate tissues. Specifically, embeding media based on EmBed 812 or LX 112 with NSA (in place of DDSA) and DMAE (replacing DMP-30), with NMA remaining constant, are formulated and offered as alternatives for routine biological work.Individual epoxy resins (Table I) or complete embedding media (Tables II-III) were tested for flow rate and viscosity. The novel media were further examined for their ability to infilftrate tissues, polymerize, sectioning and staining character, as well as strength and stability to the electron beam and column vacuum. For physical comparisons, a volume (9 ml) of either resin or media was aspirated into a capillary viscocimeter oriented vertically. The material was then allowed to flow out freely under the influence of gravity and the flow time necessary for the volume to exit was recored (Col B,C; Tables). In addition, the volume flow rate (ml flowing/second; Col D, Tables) was measured. Viscosity (n) could then be determined by using the Hagen-Poiseville relation for laminar flow, n = c.p/Q, where c = a geometric constant from an instrument calibration with water, p = mass density, and Q = volume flow rate. Mass weight and density of the materials were determined as well (Col F,G; Tables). Infiltration schedules utilized were short (1/2 hr 1:1, 3 hrs full resin), intermediate (1/2 hr 1:1, 6 hrs full resin) , or long (1/2 hr 1:1, 6 hrs full resin) in total time. Polymerization schedules ranging from 15 hrs (overnight) through 24, 36, or 48 hrs were tested. Sections demonstrating gold interference colors were collected on unsupported 200- 300 mesh grids and stained sequentially with uranyl acetate and lead citrate.

Outcome of Buccal Mucosal Graft Urethroplasty in Long Segment Anterior Urethral Stricture

2020 ◽  
Vol 19 (2) ◽  
pp. 64-68
Author(s):  
Mrinmoy Biswas ◽  
Sudip Das Gupta ◽  
Mohammed Mizanur Rahman ◽  
Sharif Mohammad Wasimuddin
Keyword(s):  
Urethral Stricture ◽  
Flow Rate ◽  
Surgical Outcome ◽  
Simple Technique ◽  
Anterior Segment ◽  
Maximum Flow ◽  
Anastomotic Site ◽  
Buccal Mucosal Graft ◽  
Male Patients

Objective: To assess the success of BMG urethroplasty in long segment anterior urethral stricture. Method: From January 2014 to December 2015, twenty male patients with long anterior segment urethral stricture were managed by BMG urethroplasty. After voiding trial they were followed up at 3 month with Uroflowmetry, RGU & MCU and PVR measurement by USG. Patients were further followed up with Uroflowmetry and PVR at 6 months interval.Successful outcome was defined as normal voiding with a maximum flow rate >15ml /sec and PVR<50 ml with consideration of maximum one attempt of OIU after catheter removal. Results: Mean stricture length was 5.2 cm (range 3-9 cm) and mean follow-up was 15.55 months (range 6-23 months). Only two patients developed stricture at proximal anastomotic site during follow-up. One of them voided normally after single attempt of OIU. Other one required second attempt of OIU and was considered as failure (5%). Conclusion: BMG urethroplasty is a simple technique with good surgical outcome. Bangladesh Journal of Urology, Vol. 19, No. 2, July 2016 p.64-68

Research on an axial-mounted dual magnetostrictive material rods-based electro-hydrostatic actuator

2021 ◽  
pp. 1045389X2110145
Author(s):  
Yuchuan Zhu ◽  
Chang Liu ◽  
Yunze Song ◽  
Long Chen ◽  
Yulei Jiang ◽  
...  
Keyword(s):  
Mathematical Model ◽  
Flow Rate ◽  
Flow Distribution ◽  
Maximum Flow ◽  
Magnetostrictive Material ◽  
Simulation Techniques ◽  
Motion State ◽  
New Type ◽  
Output Characteristics ◽  
Active Rectification

In this paper, an electro-hydrostatic actuator driven by dual axial-mounted magnetostrictive material rods-based pumps (MMPs) with a new type of active rectification valve is designed in the current study. Based on flow distribution of the active rectification valve and driving energy provided by two MMPs, the actuator can output continuous and bidirectional displacement. By establishing a mathematical model of the actuating system, using simulation techniques, the change rule of hydraulic cylinder’s motion state caused by different driving signals are studied and analyzed. Test equipment platform is constructed in the laboratory to test the output characteristics and confirm the feasibility of the new concept. The experimental results indicate that the maximum flow rate can reach approximately 2.7 L·min−1, while the operating frequency is 180 Hz.

Lichen sclerosis–induced long segment anterior urethral stricture: The early outcome of one-stage repair using dorsolateral onlay buccal mucosa graft

2021 ◽  
pp. 039156032110033
Author(s):  
Atef Fathi ◽  
Omar Mohamed ◽  
Osama Mahmoud ◽  
Gamal A Alsagheer ◽  
Ahmed M Reyad ◽  
...  
Keyword(s):  
Success Rate ◽  
Urethral Stricture ◽  
Flow Rate ◽  
Urine Volume ◽  
Maximum Flow ◽  
Maximum Flow Rate ◽  
P Value ◽  
Onlay Graft ◽  
Voiding Symptoms ◽  
Stricture Recurrence

Background: Substitution urethroplasty using buccal mucosal grafts can be performed by several approaches including ventral onlay graft, dorsal onlay graft, or ventral urethrotomy with dorsal inlay graft. Our study aims to evaluate the surgical outcome of dorsolateral buccal mucosal graft for long segment anterior urethral stricture >6 cm in patients with Lichen sclerosus (LS). Methods: A retrospective study included patients who underwent repair for long segment anterior urethral stricture >6 cm due to LS between January 2013 and April 2019. All patients were followed-up at 3, 6, 9, and 12 months postoperatively and then yearly by clinical symptoms, uroflowmetry, and calculation of post-void residual urine volume. Retrograde urethrogram was requested for patients with voiding symptoms or decreased maximum flow rate. Stricture recurrence that required subsequent urethrotomy or urethroplasty was considered failure. The success rate and surgical complications were collected and analyzed. Results: Thirty patients were identified. The median age (range) was 39 (25–61) years and a median (range) stricture length was 8 (6–14) cm. Most of postoperative complications were of minor degree. The success rate at median follow-up of 15 (12–24) months was 86.5%. The median maximum flow rate increased significantly from 6 (2–11) ml/s preoperatively to 18 (range: 6–23) ml/s at the 6th month ( p value < 0.001). Conclusion: Dorsolateral buccal mucosal grafts urethroplasty for long anterior urethral stricture caused by LS has a high success rate and low risk of complications including stricture recurrence.

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