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PubMed2001

New insights into the applicability of T-cell receptor gamma gene rearrangement analysis in cutaneous T-cell lymphoma.

BACKGROUND: Detection of clonal T-cell receptor (TCR) gamma gene rearrangement by polymerase chain reaction (PCR) based method is a marker for cutaneous T-cell lymphoma (CTCL) although it can be seen in some benign dermatoses. To determine the accuracy of histologic criteria alone as well as the adjuvant diagnostic role of TCR gene rearrangement for the diagnosis of CTCL, we studied 100 patients with cutaneous T-cell infiltrates by both histology and TCR gene rearrangement. METHODS: The histologic features of the 100 patients were first reviewed by two independent dermatopathologists and their confidence in the diagnosis of CTCL was assigned one of four levels. Then the specimens were analyzed for TCR gene rearrangement either on paraffin-embedded or fresh-frozen tissue by PCR/denaturing gradient gel electrophoresis (DGGE). RESULTS: The clonality was detected in 100% (15/15) diagnostic of, 84.6% (11/13) consistent with, 57.6% (19/33) suggestive of CTCL. In 9 cases TCR gene rearrangement was compared between formalin-fixed and fresh specimens of the same individual, but with different degrees of histologic confidence (no lower than suggestive). In all cases fresh specimens were positive. In 5 of the cases (2-diagnostic, 2-consistent, 1-suggestive) formalin-fixed specimens were positive as well, and in 4 cases (1-consistent, 3-suggestive) formalin-fixed specimens were negative. When TCR gene rearrangement was studied in eight cases on sequential biopsies from the same patient, the clonality was detected in only one or two biopsies in four cases in which the histologic confidence was low (suggestive or nondiagnostic). The TCR gene rearrangement study showed identical banding patterns in lesions from different clinical stages in most patients. However, we observed that in one case, oligoclonal-banding pattern was seen in initial biopsy with histopathologic consistent with CTCL, while monoclonal banding pattern in more advanced lesion. CONCLUSIONS: Our data have demonstrated that TCR gene rearrangement studies by PCR/DGGE are consistently positive regardless of tissue fixation (formalin-fixed, paraffin-embedded vs. fresh-frozen tissue) and biopsy site when the histologic degree of confidence is very high (diagnostic). So, it may be of less importance as an adjuvant to histopathologic diagnosis for the cases with diagnostic CTCL histology. However, TCR gene rearrangement studies are particularly important in earlier cases with less conclusive histology, which provides strong confirmatory evidence of an evolving CTCL. In these cases, multiple biopsies may be required to establish the diagnosis and analysis of fresh tissue is suggested to increases the sensitivity. Moreover, our observation also suggested that some CTCL might not be monoclonal de novo, but oligoclonal instead.

Clone Cells↗
PubMed1990

Immunolabeling of grapevine flavescence dorée MLO in salivary glands of Euscelidius variegatus: a light and electron microscopy study.

Flavescence dorée (FD), a grapevine yellows disease, is caused by a mycoplasma-like organism (MLO). A colloidal gold indirect immunolabeling technique identified MLO in salivary glands of a vector leafhopper, Euscelidius variegatus. After aldehyde fixation, tissue samples were prepared by cryoultramicrotomy or embedding in acrylic resins. Double fixation with aldehydes and osmium retroxide, followed by embedding in epon, was also performed. Thin or semi-thin serial sections were treated with polyclonal anti-FD-MLO rabbit antibodies, then with gold-conjugated anti-rabbit IgG. Labeling was revealed using the silver enhancement technique for light microscopy. MLO in frozen thin sections of glands were efficiently labeled. Optimal results were obtained with 4% paraformaldehyde-0.1% glutaraldehyde fixation and low-temperature embedding in LR White resin. Both scattered MLO and unusual dense forms of MLO were easily detected with the electron-dense gold probe. This method distinguished MLO from other membrane-limited bodies and provided a good tool for studying infection in large regions of FD-infected tissues by light microscopy.

Animals↗
PubMed1993

Natural preload of aortic valve leaflet components during glutaraldehyde fixation: effects on tissue mechanics.

The mechanics of glutaraldehyde-fixed aortic valve leaflets depend largely on the amount of stress present during fixation. Our previous work has suggested that even when the aortic valve is flaccid, the leaflet components are preloaded. We have, therefore, hypothesized that fixing valve leaflets in this naturally preloaded state will affect the function of their components, the fibrosa and the ventricularis. We have compared the elastic response of fibrosa and ventricularis fixed under 'low' and 'zero' tensile and compressive preload by testing 120 of these layers: (i) fresh, (ii) glutaraldehyde-fixed, and (iii) isolated from whole porcine aortic valve leaflets fixed while intact. In both the radial and circumferential directions, the fibrosa from intact-fixed valves was more extensible than the fresh (39.2 vs 29.2% strain to high modulus phase at p < 0.0122, and 12.7 vs 8.1% strain, at p < 0.0003, respectively). The ventricularis from intact-fixed valves, however, was less extensible than when fresh (35.4 vs 63.7% strain, at p < 0.00001 in the radial direction). The fibrosa must have, therefore, been fixed under compression and the ventricularis under tension, when fixed together in the intact aortic valve cusp. The tensile stresses in the intact-fixed ventricularis produced a greater circumferential elastic modulus than in separately fixed tissue (9.62 vs 4.65 MPa, at p < 0.00001), likely through a fibre recruitment process. Compressive stresses in the fibrosa produced a decrease in the elastic modulus both radially and circumferentially (from 3.79 to 2.26 MPa at p < 0.0023, and from 9.55 to 4.65 MPa at p < 0.00001, respectively). Fixing porcine aortic valves at even minimal tensile and compressive preload, such as that which occurs naturally, significantly alters both the extensibility and the elastic modulus of the valve leaflet components.

Animals↗
PubMed1998

Assessment of glutaraldehyde crosslinking efficiency with an amine-specific fluorescent probe.

BACKGROUND: Crosslinking of heart valves with glutaraldehyde involves the binding of amine groups. We have developed a technique that provides an inverse measure of the degree of tissue fixation by quantifying the amount of unbound amines. METHODS: Whole aortic valves were exposed to 0.5% glutaraldehyde solution for 0, 1, 15, and 60 minutes, 6 hours, and 1 and 7 days. Frozen sections were exposed to carboxyfluorescein succinimidyl ester, a fluorescent amine-reactive probe. Images were acquired from each section and processed to separate pixels representing tissue from those representing background. An average fluorescent intensity for each image was calculated and related to the number of unbound amines by comparing with standards. RESULTS: The amount of uncrosslinked amines was observed to decrease exponentially with fixation time and achieved a plateau at 1 day of fixation. A significant difference in the amount of unbound amines also exists between valve leaflets fixed while connected to the root and those excised from the root before fixation. CONCLUSIONS: This amine measurement technique, being sensitive to spatially varying differences in chemical fixation, should be useful in evaluating the efficacy of new fixation protocols.

Amines↗
PubMed1997

In situ hybridization: identification of rare mRNAs in human tissues.

In situ hybridization is used for detection of RNA expression when conservation of tissue architecture is important. Most in situ hybridization protocols are written for tissues from animals (i.e., rat) which can be harvested and preserved rapidly. In contrast, human tissue is more difficult to obtain, hence in situ hybridization experiments must frequently be performed with less than optimal tissue preservation. This procedure details hybridization of a radiolabeled single-stranded RNA probe (riboprobe) to complementary sequences of cellular RNA in human tissue sections. This method enables detection of rare mRNA species in specific cell types of human tissue, offering distinct advantages over other in situ methods due to increased sensitivity. In particular, we have found that UV cross-linking and ribonuclease treatment protocols need to be altered for human tissues to ensure successful results, making this protocol unique to those previously described. In situ hybridization experiments can be performed using either DNA or RNA probes. RNA probes are advantageous since they form stable hybrids, are single-stranded, have little or no reannealing during hybridization, and can be synthesized to high specific activity. RNA probes can be readily created utilizing SP6, T3, or T7 promoters in both sense and antisense orientations to provide non-specific (control) and specific probes. Disadvantages of RNA riboprobes include a tendency for RNA to stick non-selectively more than DNA, and degradation by RNase (hence strict adherence to RNase-free precautions is mandatory during most of the protocol). The following protocol includes: (1) preparation of human tissues (tissue fixation and sectioning are highlighted as critical for probe penetration, preservation of tissue architecture, retention of tissue RNA, and overall success); (2) generation of radiolabeled riboprobes (total incorporation of radionucleotide is important to increase sensitivity; 35S was chosen as a compromise between excellent sensitivity, cellular resolution, and required exposure times (compared with 32P or 3H); non-isotopic methods have not been tested in a side-by-side comparison with 35S in human tissues by us, but theoretically might offer faster exposure times while maintaining high resolution); (3) hybridization conditions (stringency, temperature, washes, tissue dehydration); and (4) sample visualization (application of photographic emulsion, developing, fixing, staining, and counterstaining of individual slides).

Humans↗
PubMed1984

Red cell fragmentation in human disease (a light and scanning electron microscope study).

Although the mechanism of schizocyte formation has been amply documented in animal experiments and in in-vitro models, the fragmentation encounter between flowing red cells and fibrin strands has not previously been successfully demonstrated in human, microangiopathic disease. If blood flow is restored briefly in vitro immediately prior to tissue fixation, the instant of red cell fragmentation can be captured. Examination of tissue specimens fixed in this manner shows a pathophysiologic process that amplifies the findings previously described in other studies. In the patient under study, the microangiopathic process was widespread in all specimens of pulmonary and renal tissue that had been fixed after brief restoration of blood flow. Small arteries as well as the true microcirculation of both organs were involved. The microangiopathic process in the small arteries and arterioles presented as a partially occlusive thrombus of characteristic histology. The pulmonary capillaries contained linear fibrin microclots festooned with distorted and partially fragmented red cells. The microcirculation of the kidney showed the same findings as well as amorphous, sludged, occlusive red cell masses, particularly in the renal medulla.

Adenocarcinoma↗
PubMed1980

Cytochemical localization of adenylate cyclase.

This study examined 1) the effect of lead and fixation on adenylate cyclase activity, 2) the effect of lead on App(NH)p, and 3) the specificity of App(NH)p as a substrate for adenylate cyclase under the conditions of the cytochemical assay. The results indicated that: 1) fixation that provides adequate structural preservation inhibits enzyme activity to varying degrees depending on the tissue, fixative, length and temperature of fixation; 2) millimolar concentrations of lead do not negatively affect the adenylate cyclase activity of several different tissues (especially if 10 mM NaF is present); 3) lead does not cause the nonenzymatic hydrolysis of App(NH)p; 4) the App(NH)p obtained from the supplier is contaminated and should be purified before use, since lead can interact with the contaminants and this may be a source of error in the assay; and 5) adenylate cyclase appears to be the major enzyme that cleaves App(NH)p under cytochemical conditions.

Adenylyl Cyclases↗
PubMed1985

Management of infected fractures of the tibia with associated soft tissue loss: experience with external fixation, bone grafting and soft tissue reconstruction using pedicle muscle flaps or microvascular composite tissue grafts.

The aim of this study was to assess the results of treatment given by a team of orthopaedic and plastic surgeons in a series of infected unhealed fractures of the tibia associated with loss of adjacent soft tissues. Twenty-five lower leg fractures, treated during a 10 year period, entered the study and were grouped according to the principles of treatment followed. In the earlier Group A (nine patients), the osteosynthesis implants were retained or changed to more stable internal fixation devices, the soft tissue defects were closed by conventional muscle or musculocutaneous flaps and bone grafting procedures were performed late in the treatment scheme. In the later Group B (16 patients) the implants were removed and the fracture stabilised by external fixation; the defects were covered with pedicle muscle flaps or with microvascular composite tissue grafts and cancellous bone grafting was performed at the same operation. Twenty-three fractures healed. One fracture developed non-union and in one patient infection necessitated below-knee amputation. The time of union after surgical reconstruction was significantly shorter in Group B (24 +/- 3 weeks) than in Group A (47 +/- 11 weeks). The results suggested that: in severe infected fractures of the tibia surgical implants used previously for fracture treatment should be removed and replaced with an external frame using firm axial compression, microvascular composite grafts seem to improve greatly the rate of healing, early bone grafting should be included in the reconstruction and late infections can be largely avoided even after extensive one-stage reconstructive procedures.

Adolescent↗
PubMed2004

In vitro comparison of standard and Knotless metal suture anchors.

PURPOSE: Clinical experience after failed Knotless suture anchor (Mitek, Westwood, MA) fixations suggested that the Knotless anchor provides considerably less fixation stability than a standard metal anchor. The purpose of this study was to analyze soft tissue fixation to bone comparing a standard and a Knotless metal suture anchor. TYPE OF STUDY: In vitro study. METHODS: The Mitek GII and Mitek Knotless suture anchors were tested on 7 human cadaveric fresh-frozen glenoids. The anchors were inserted into the glenoid rims, and the sutures of the anchors were fixed to a metal hook attached to the cross-head of a testing machine. Cyclic loading was performed. The gap formation between the metal hook and the glenoid rim, the ultimate failure loads and the modes of failure were determined. RESULTS: The mean gap formation was significantly greater for the Knotless anchor (3.8 +/- 1.4 mm) than for the GII anchor (2.4 +/- 0.5 mm) after 25 cycles with 50 N repeated load (P =.04). The largest gap of a Knotless fixation was 5.3 mm compared with 3.0 mm for the GII. The ultimate failure load was not significantly different for the Knotless anchor (179 N) and for the GII anchor (129 N). Both anchors failed by either rupture of the suture material or by pullout of the anchors. CONCLUSIONS: The GII anchor allows significantly less displacement than the Knotless anchor. Ultimate tensile strength and mode of failure are similar. Greater displacement results in larger gap formation between the soft tissue and the bone. This might weaken and jeopardize the repair. CLINICAL RELEVANCE: If reattached soft tissues are subjected to postoperative loading, gap formation may result when using the Knotless anchor. For these conditions, suture fixation with knots may be used instead.

Equipment Design↗
PubMed2002

Isolation of total-RNA from formalin-fixed rat retina.

The aim of this project was to establish a method for the purification of total-RNA from fixed rat-retina. Two different established methods were used for RNA purification, and successful isolation was verified with RT-PCR for amplification of beta-actin (two different product-lengths) and subsequent gel-electrophoresis. Total-RNA was successfully isolated from fixed rat-retina. The house keeping gene, beta-actin could be detected after fixing the retina either with 1% formalin or with 4% paraformaldehyde (PFA). Hexamer-primer based RT-PCR gave better results than the oligo-d(T)-primer based RT-PCR method. Both the 698 and 225 bp beta-actin-fragments could be successfully amplified, where amplification of the latter was more efficient. This approach shows that tissue fixation prior to RNA-isolation facilitates the rapid isolation of undamaged RNAs in tissues such as the retina, which are known to yield low levels of RNA and are vulnerable to RNases.

Animals↗
PubMed1998

Characterization of cytoplasmic secretory granules (PSG), in prostatic epithelium and their transformation-induced loss in dysplasia and adenocarcinoma.

Cytoplasmic clarity is a histological feature of normal prostatic secretory cells, but in this study, tissue fixation in strong (>2.5%) glutaraldehyde dramatically altered cytological staining. Secretory cytoplasm appeared red and granular on routine stains because of myriad intensely staining eosinophilic granules (PSG). Immunostaining for prostate-specific antigen (PSA) and prostatic acid phosphatase (PAP) showed their exclusive localization to the PSG. Electron microscopy confirmed these findings and also showed that after fixation in many agents, including formaldehyde, PSG appeared empty, accounting for the artefactual "clear cell" appearance on light microscopy. PSG were most densely concentrated apically in a bud-shaped luminal compartment in which cytokeratin was selectively absent. Normal exocrine secretion was visualized as detachment of apocrine buds or their in situ disintegration. Distinctively in dysplasia and almost all carcinomas, PSG were rare to absent, and proteases were free in the cytoplasm, often concentrated beneath the apical membrane. The apocrine compartment was absent, with no observed secretory mechanism. Tumor cells had dark amphiphilic cytoplasm after all fixatives. This provided a reliable method of distinguishing malignant from benign glands in tissues fixed in strong glutaraldehyde. Clear cell carcinomas, whose cytoplasm mimicked routinely fixed normal secretory cells, surprisingly had almost no PSG. Instead, their "granules" were lipid-filled vacuoles reflecting a secretory pathway not seen in normal cells, dysplasia, or the common "dark cell" carcinomas. These observations may define two distinctive biological pathways of prostate cancer evolution and may facilitate diagnostic decisions on needle biopsy samples.

Acid Phosphatase↗
PubMed2001

Effect of fixation, soft-tissues, and scan projection on bone mineral measurements with dual energy X-ray absorptiometry (DXA).

The objective of this study was to determine the effect of fixation, soft tissues, and scan projection on bone mineral measurements with dual energy X-ray absorptiometry (DXA). In seven fresh cadavers, DXA scans were obtained within 48 hours of death and after 10 months of fixation with 5% formalin/95% ethanol. The measurements showed a high linear relationship (r2 > 0.97; SEE% < 10%), with no significant deviation after fixation (except for total body BMD: -3.1%). In 14 specimens, the precision of femoral and spinal analyses was determined under in situ and ex situ conditions. There was no significant difference between ex situ and in situ reproducibility, the coefficient of variation being < 3% for the BMC and < 2% for the BMD (except at the greater trochanter). The effect of the soft tissues and scan projection was assessed in 83 cadavers aged 80.4 +/- 10.3 years. The soft tissues had only a small effect on analyses of the total femur (r2 > 0.90; SEE% < 9%), but led to more substantial deviations in regional femoral analyses and in the spine (r2 = 0.78-0.90; SEE% = 8-22%). Comparing lateral with anterior-posterior (AP) spinal scans, the vertebral bodies were found to occupy 40.2 +/- 7.2% of the BMC, and 62.0 +/- 11.2% of the BMD, the ranges being 26-58%, and 38-91%, respectively. There were large deviations from linearity between in situ AP and ex situ lateral spinal scans with r2 values of 0.63 and 0.73 for BMD and BMC (SEE% = 52% and 27% relative to the vertebral body), respectively.

Absorptiometry, Photon↗
PubMed1995

Glutathione S-transferases and gamma-glutamyl transpeptidase in the rat nervous systems: a basis for differential susceptibility to neurotoxicants.

Glutathione and its related enzymes play a major role in the detoxification of toxic chemicals. In rat brain the pattern of distribution of reduced glutathione exhibits cellular heterogeneity, suggesting also the possibility of cellular differences in glutathione conjugating capacity. To understand the potential role of GSH in detoxification of neurotoxicants, the distributions of the glutathione conjugating and metabolizing enzymes, glutathione S-transferase (GST; alpha-, mu- and pi-classes) and gamma-glutamyl transpeptidase (gamma-GT) were determined immunohistochemically in brain, lumbar spinal cord and dorsal root ganglia (DRG) of adult Sprague-Dawley rats using polyclonal antibodies. The influence of tissue fixation on apparent distribution was also examined. Glial cells and neurons throughout the nervous system were only weakly positive with alpha-GST in frozen sections. No immunoreactivity for the alpha-class GSTs was observed in any of the paraformaldehyde-fixed neural specimens examined. In microwave-fixed frozen sections, immunoreactivity to mu-GST was found in astrocytes and neurons throughout the brain and spinal cord, and in the neurons and satellite cells of the DRG. Immunoreactivity for pi-GST was seen in oligodendrocytes but not in astrocytes in any region of the CNS examined. Similarly, satellite cells of the DRG were positive for pi-GST. Neuronal perikarya of the entire neopallium, hippocampus, cerebellum, brainstem, spinal cord and DRG were also positively stained for pi-GST. The differential staining of astrocytes and oligodendrocytes with pi- and mu-GST was unaltered in paraformaldehyde fixed tissues, but the neuronal immunostaining was lost. The ependyma, pia and choroid plexus stained positively with all three GST antibodies regardless of fixation. Gamma-Glutamyl transpeptidase-like immunoreactivity was confined to non-neuronal elements of both central and peripheral nervous systems. Ependymal cells throughout the central nervous systems stained intensely with antibodies directed against gamma-GT. Satellite and Schwann cells of the DRG and glial cells of the spinal cord and brain exhibited moderate to intense immunoreactivity for gamma-GT. The heterogeneous cellular distribution of glutathione and its metabolizing enzymes may reflect cellular differences in capacity for metabolic processing of both endogenous compound and xenobiotics.

3,3'-Diaminobenzidine↗
PubMed2004

Interference screw fixation of soft tissue grafts in anterior cruciate ligament reconstruction: part 1: effect of tunnel compaction by serial dilators versus extraction drilling on the initial fixation strength.

BACKGROUND: Compaction of the bone-tunnel walls by serial dilation is believed to enhance the interference screw fixation strength of the soft tissue grafts in anterior cruciate ligament (ACL) reconstruction. HYPOTHESIS: Serial dilation enhances the fixation strength of soft tissue grafts in ACL reconstruction over extraction drilling. STUDY DESIGN: Randomized experimental study. METHODS: Initial fixation strength of the doubled anterior tibialis tendon grafts (fixed with a bioabsorbable interference screw) was assessed in 21 pairs of human cadaver tibiae with either serially dilated or extraction-drilled bone tunnels. The specimens were subjected to a cyclic-loading test, and those surviving were then tested using the single-cycle load-to-failure test. RESULTS: During the cyclic-loading test, there were 3 fixation failures in the serially dilated and 6 failures in the extraction-drilled specimens but no significant stiffness or displacement differences between the groups. In the subsequent load-to-failure test, the average yield loads were 473 +/- 110 N and 480 +/- 115 N for the 2 groups respectively (P =.97) and no difference with regard to stiffness or mode of failure. CONCLUSIONS: Serial dilation does not increase the strength of interference fixation of soft tissue grafts in ACL reconstruction over extraction drilling. CLINICAL RELEVANCE: The results of this experiment do not support the use of serial dilators in ACL reconstruction.

Adolescent↗
PubMedDate not supplied

Applications of immunogold-silver enhancement: testing of monoclonal antibodies and detection of human cytomegalovirus in histologic specimens.

Human cytomegalovirus (HCMV) is an important pathogen in neonates, transplant patients, and individuals with acquired immunodeficiency syndrome (AIDS). Reliable techniques for the detection of this virus in clinical specimens would aid in improving methods for diagnosis and increasing our understanding of viral pathogenesis. We evaluated the utility of immunogold-silver enhancement to determine the specificity of murine monoclonal antibodies directed against HCMV proteins and applied these antibodies to the detection of HCMV in histologic material. Nine antibodies were tested in a tissue-culture system to determine the location of staining. All were found to be active in frozen tissue, but only two of these antibodies were reactive in formalin-fixed tissue. We also evaluated a novel tissue fixative technique (AMeX; Fixation and dehydration with acetone) that has been used to maintain antigenicity of T-lymphocyte cell markers. All antibodies remained reactive against their respective HCMV proteins in tissue fixed by this technique. However, dehydration of tissue may limit the usefulness of AMeX fixation. Immunogold-silver enhancement is a useful and reliable histochemical technique for detection of HCMV antigens in pathologic specimens.

Antibodies, Monoclonal↗
PubMed2002

Cyanoacrylate tissue adhesives for skin closure in the outpatient setting.

Cyanoacrylate and fibrin tissue adhesives can aid the facial plastic surgeon performing surgery in the outpatient setting. Octyl-2-cyanoacrylate is ideal for superficial skin closure and eliminates the need for suture removal. Subcutaneous sutures are needed in most cases to provide the best cosmetic outcomes. Fibrin tissue adhesives can be used to aid in attaining hemostasis or to seal or fixate tissues. Cyanoacrylate and fibrin-based adhesives can simplify office-based facial plastic surgery.

Ambulatory Care↗
PubMed1988

Variability of fiber size, capillary density and capillary length related to horse muscle fixation procedures.

Muscle samples were taken from the superficial part of the semitendinosus muscle of four standardbred horses to compare the effects of three different fixation procedures on fiber size and capillarity. Infiltration fixation in an extended position of the muscle was compared to immersion fixation and infiltration fixation in a flexed position of the limb. The capillary length density was used to estimate muscle capillarity. Infiltration fixation was found to be a viable alternative to perfusion fixation of muscles for morphometry of fiber size and capillary length density allowing muscle tissue fixation at a defined sarcomere length without fiber kinking.

Animals↗
PubMed1995

Combining microwave stabilization and microwave-stimulated fixation of brain tissue with microwave-stimulated staining.

Microwave stabilization of rat brain tissue using saline was compared with microwave-stimulated fixation using either formaldehyde or Kryofix. Sections were stained according to the microwave-adapted Nissl, Klüver-Barrera, and Bodian methods. Depending on the preparative choice the microwave staining method had to be adjusted. Optimal combinations of preparative techniques and staining methods are feasible. In most cases the microwave saline-stabilized and microwave Kryofix-treated sections were better suited for application in neurohistology than the microwave formaldehyde-treated sections for light microscopy.

Animals↗