摘要:

We report a modification of the immunogold-silver staining method (IGSS) for localizing hepatic phosphoenolpyruvate carboxykinase (PEPCK) in tissue sections, and we compare the efficacy of localizing the primary antibody with either a 5 nm gold labeled secondary antibody or 5 nm gold labeled secondary and tertiary antibodies. Light microscope examination of 10μm frozen sections demonstrated that the use of combined secondary and tertiary gold labeled antibodies was superior to using a secondary gold labeled antibody alone. The increased labeling density (number of colloidal gold particles/antigenic site/cell) achieved by combined gold labeled antibodies was confirmed by electron microscopy. The increased labeling density resulted in a two-thirds reduction in the time needed for the IGSS physical development of the silver shells and less background. We achieved intense specific staining of hepatocytes expressing PEPCK while minimizing background staining. The use of combined secondary and tertiary gold labeled antibodies enhances the signal-to-noise ratio, achieves high resolution and is a suitable method for use in both light and electron microscopy.

ISSN:1052-0295    

DOI:10.3109/10520299309105636

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

A histological method is described for determining the proportion of growing hair follicles h skin samples. A variation of the Sacpic staining method, modified for bulk processing, produces high contrast staining of the principal tissue types present in skin. In particular, the inner root sheath is accentuated, facilitating detection of active follicles. Skin preparations from a range of species are used to illustrate structural characteristics of follicles viewed in cross section at various stages of the hair cycle and to establish criteria for classification of the state of activity of follicles. The hair cycle may be divided into quiescent and active states at the points of rapid transition (early pronanagen and mid catagen). Data from repeated skin biopsies from ferrets and goats are also used to demonstrate quantitative estimation of follicle activity, change in compound follicle size, and the relationship between follicle type and fiber medullation.

ISSN:1052-0295    

DOI:10.3109/10520299309105637

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

As an approach to the study of inflammation, strategies were evaluated for quantitative assessment of plasma exudate from thecutaneous microvasculature. Measurements were based on recovery of Evans blue dye (EB) from rat skin. After preliminary studies to evaluate extraction methods, almost complete EB recovery was accomplished by homogenizing tissue in a mixture of acetone, water and sodium sulfate. When sources of potential variation were identified, expression of results as agonist-induced plasma accumulation provided precise results based on EB measurements. Also. the feasibility of parallel biochemical studies was demonstrated.

ISSN:1052-0295    

DOI:10.3109/10520299309105638

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

Orthopedic biomaterials currently are made of metal alloy coated with one or more thin layers of dense or porous ceramic or metal. Sections of these materials implanted in human bone were made without altering the implant or bone-implant interfaces. Bone containing an implant was fixed and then embedded in polyme-thylmethacrylate. Thick sections were made using a cooled. low speed diamond saw, then ground and polished. Some were stained by fuchsin-toluidine staining solution, others were acid etched to reveal the structure of the metal contained in the prosthesis. Observation by reflected and transmitted light microscopy revealed microstructure of the implant material as well as features of the surrounding tissues.

ISSN:1052-0295    

DOI:10.3109/10520299309105639

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

As an approach to the study of inflammation, strategies were evaluated for quantitative assessment of plasma exudate from the cutaneous microvasculature. Measurements were based on recovery of Evans blue dye (EB) from rat skin. After preliminary studies to evaluate extraction methods, almost complete EB recovery was accomplished by homogenizing tissue in a mixture of acetone, water and sodium sulfate. When sources of potential variation were identified, expression of results as agonist-induced plasma accumulation provided precise results based on EB measurements. Also, the feasibility of parallel biochemical studies was demonstrated.

ISSN:1052-0295    

DOI:10.3109/10520299309105640

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

4′,6-Diamidino-2-phenyl-indole (DAPI), is a fluorescent probe that specifically and quantitatively stains DNA. Electroporation of viablePetuniaprotoplasts in the presence of DAPI revealed integral fluorescence that was similar for both the electroporated and fixed protoplasts. indicating quantitative staining of DNA. DAPI fluorescence was localized in the nuclei of viable protoplasts ofPetunia. Protoplasts had a short term viability of 56–65% of the control (non-electroporated. unstained) protoplasts as determined by fluorescein diacetate staining 24 hr following electroporation in the presence of DAPI. The majority (84% of the number originally cultured) of these protoplasts subjected to electroporation were able to form a cell wall, but most did not form microcalli because they were blocked in cell division. The three week plating efficiency for protoplasts exposed to DAPI was 4% of the original number of protoplasts initially cultured compared to 30% for the control. DAPI should not be used as a fluorescent probe for plant protoplasts when the protoplasts are cultured for sustained growth because the levels of DAPI required to obtain quantitative staining of the DNA resulted in inhibition of the cell cycle. DAPI may, however, be used as a fluorescent DNA probe for short term (24 hr) studies.

ISSN:1052-0295    

DOI:10.3109/10520299309105641

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

Agarose was used to embed the brain or spinal cord of lampreys or rats before cutting vibratome sections. Agarose embedding was compatible with immunocytochemistry or the use of horseradish peroxidase as a neuroanatomical tracer. Concentrated agarose with high intrinsic gel strength was optimal for embedding glutaraldehyde fixed neural tissue. A quick procedure was to blot tissue and embed in 5% (w/v] Sigma type I-A or Litex type LSL agarose at 45–55 C dissolved in 50 mM neutral-pH TFUS buffer before cutting 50–100μmvibratome sections. An alternative procedure that improved retention of tissue sections in the agarose was to rinse the tissue in H20, blot and embed in 5% (w/v] Sigma type I-A or Litex type LSL agarose at 45–55 C dissolved in H20, then equilibrate the block overnight in buffer. Phosphate buffer prevented complete dissolving of agarose. Tissue could be covalently linked to the embedding matrix using a novel aldehyde-derived agarose (NuFix®FMC BioProducts). Slices of spinal cord from neonatal rats could be cut after embedding in 5% FMC Seaprep®agarose in rat Ringer's at 23–26 C.

ISSN:1052-0295    

DOI:10.3109/10520299309105642

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

ISSN:1052-0295    

DOI:10.3109/10520299309105643

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

ISSN:1052-0295    

DOI:10.3109/10520299309105644

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor