Preservation of Subcellular Structures via the Combined Langmuir Trough-Critical Point Drying Technique

1976 ◽  
Vol 95 (4) ◽  
pp. 702 ◽  
Author(s):  
Glenn K. Walker ◽  
Timothy K. Maugel ◽  
Dennis Goode
Keyword(s):  
Critical Point ◽  
Langmuir Trough ◽  
Critical Point Drying

scholarly journals THE HUMAN CHROMOSOME

1969 ◽  
Vol 41 (1) ◽  
pp. 73-90 ◽  
Author(s):  
J. G. Abuelo ◽  
Dorothy E. Moore
Keyword(s):  
Tissue Culture ◽  
Electron Microscope ◽  
Human Chromosome ◽  
Critical Point ◽  
Human Lymphocytes ◽  
Langmuir Trough ◽  
Trypsin Treatment ◽  
Short Term ◽  
Dnase Digestion ◽  
Critical Point Drying

Human lymphocytes were grown in short-term tissue culture and were arrested in metaphase with Colcemid. Their chromosomes were prepared by the Langmuir trough-critical point drying technique and were examined under the electron microscope. In addition, some chromosomes were digested with trypsin, Pronase, or DNase. The chromosomes consist entirely of tightly packed, 240 ± 50-A chromatin fibers. Trypsin and Pronase treatments induce relaxation of fiber packing and reveal certain underlying fiber arrangements. Furthermore, trypsin treatment demonstrates that the chromatin fiber has a 25–50 A trypsin-resistant core surrounded by a trypsin-sensitive sheath. DNase digestion suggests that this core contains DNA.

The Critical Point Drying Method Applied to Scanning Electron Microscopy of L-929 Cells

1970 ◽  
Vol 28 ◽  
pp. 278-279
Author(s):  
Charles TurnbiLL ◽  
Delbert E. Philpott
Keyword(s):  
Electron Microscopy ◽  
Carbon Dioxide ◽  
Surface Tension ◽  
Critical Point ◽  
Freeze Drying ◽  
Attractive Alternative ◽  
Crystal Formation ◽  
Critical Point Drying ◽  
Time Required ◽  
Scanning Electron

The advent of the scanning electron microscope (SCEM) has renewed interest in preparing specimens by avoiding the forces of surface tension. The present method of freeze drying by Boyde and Barger (1969) and Small and Marszalek (1969) does prevent surface tension but ice crystal formation and time required for pumping out the specimen to dryness has discouraged us. We believe an attractive alternative to freeze drying is the critical point method originated by Anderson (1951; for electron microscopy. He avoided surface tension effects during drying by first exchanging the specimen water with alcohol, amy L acetate and then with carbon dioxide. He then selected a specific temperature (36.5°C) and pressure (72 Atm.) at which carbon dioxide would pass from the liquid to the gaseous phase without the effect of surface tension This combination of temperature and, pressure is known as the "critical point" of the Liquid.

SEM Cold Stage Techniques for the Observation of Vegetative and Floral Apices of Oat (Avena sativa L.) Plants

1977 ◽  
Vol 35 ◽  
pp. 590-591
Author(s):  
B. K. Kirchoff ◽  
L.F. Allard ◽  
W.C. Bigelow
Keyword(s):  
Critical Point ◽  
Avena Sativa ◽  
Substantial Improvement ◽  
Fresh Tissue ◽  
Cold Stage ◽  
Critical Point Drying ◽  
Almost All ◽  
Cell Collapse ◽  
Conductive Paint ◽  
Carbon Based

In attempting to use the SEM to investigate the transition from the vegetative to the floral state in oat (Avena sativa L.) it was discovered that the procedures of fixation and critical point drying (CPD), and fresh tissue examination of the specimens gave unsatisfactory results. In most cases, by using these techniques, cells of the tissue were collapsed or otherwise visibly distorted. Figure 1 shows the results of fixation with 4.5% formaldehyde-gluteraldehyde followed by CPD. Almost all cellular detail has been obscured by the resulting shrinkage distortions. The larger cracks seen on the left of the picture may be due to dissection damage, rather than CPD. The results of observation of fresh tissue are seen in Fig. 2. Although there is a substantial improvement over CPD, some cell collapse still occurs.Due to these difficulties, it was decided to experiment with cold stage techniques. The specimens to be observed were dissected out and attached to the sample stub using a carbon based conductive paint in acetone.

The Effects of Drying Techniques on Clay-Rich Soil Texture

1974 ◽  
Vol 32 ◽  
pp. 466-467
Author(s):  
T. G. Naymik
Keyword(s):  
Critical Point ◽  
Liquid Nitrogen ◽  
Liquid Nitrogen Temperature ◽  
Drying Methods ◽  
Air Drying ◽  
Freeze Dried ◽  
Critical Point Drying ◽  
Set Up ◽  
The Relationship ◽  
Quartz Grains

Three techniques were incorporated for drying clay-rich specimens: air-drying, freeze-drying and critical point drying. In air-drying, the specimens were set out for several days to dry or were placed in an oven (80°F) for several hours. The freeze-dried specimens were frozen by immersion in liquid nitrogen or in isopentane at near liquid nitrogen temperature and then were immediately placed in the freeze-dry vacuum chamber. The critical point specimens were molded in agar immediately after sampling. When the agar had set up the dehydration series, water-alcohol-amyl acetate-CO2 was carried out. The objectives were to compare the fabric plasmas (clays and precipitates), fabricskeletons (quartz grains) and the relationship between them for each drying technique. The three drying methods are not only applicable to the study of treated soils, but can be incorporated into all SEM clay soil studies.

Preservation of Plant Cell Surfaces For Scanning Electron Microscopy

1973 ◽  
Vol 31 ◽  
pp. 472-473
Author(s):  
Linda M. Sicko ◽  
Thomas E. Jensen
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Critical Point ◽  
Filter Paper ◽  
Freeze Drying ◽  
Cell Surfaces ◽  
Air Drying ◽  
Popular Method ◽  
Critical Point Drying ◽  
Scanning Electron

The use of critical point drying is rapidly becoming a popular method of preparing biological samples for scanning electron microscopy. The procedure is rapid, and produces consistent results with a variety of samples. The preservation of surface details is much greater than that of air drying, and the procedure is less complicated than that of freeze drying. This paper will present results comparing conventional air-drying of plant specimens to critical point drying, both of fixed and unfixed material. The preservation of delicate structures which are easily damaged in processing and the use of filter paper as a vehicle for drying will be discussed.

Rapid Critical Point Drying of Tissues in a Parr Bomb

1972 ◽  
Vol 30 ◽  
pp. 400-401
Author(s):  
C.A. Baechler ◽  
W. C. Pitchford ◽  
J. M. Riddle ◽  
C.B. Boyd ◽  
H. Kanagawa ◽  
...  
Keyword(s):  
Critical Point ◽  
Critical Pressure ◽  
Soft Tissues ◽  
Biological Tissues ◽  
Critical Temperatures ◽  
Air Drying ◽  
The Past ◽  
Critical Point Drying ◽  
Great Stress ◽  
Infiltration Techniques

Preservation of the topographic ultrastructure of soft biological tissues for examination by scanning electron microscopy has been accomplished in the past by using lengthy epoxy infiltration techniques, or dehydration in ethanol or acetone followed by air drying. Since the former technique requires several days of preparation and the latter technique subjects the tissues to great stress during the phase change encountered during air-drying, an alternate rapid, economical, and reliable method of surface structure preservation was developed. Turnbill and Philpott had used a fluorocarbon for the critical point drying of soft tissues and indicated the advantages of working with fluids having both moderately low critical pressures as well as low critical temperatures. Freon-116 (duPont) which has a critical temperature of 19. 7 C and a critical pressure of 432 psi was used in this study.

Modified gach technique vs critical point drying: Shrinkage analysis for SEM

1987 ◽  
Vol 45 ◽  
pp. 954-955
Author(s):  
B. Thompson ◽  
N. Sculov ◽  
R.E. Crang
Keyword(s):  
Critical Point ◽  
Calf Serum ◽  
Drying Shrinkage ◽  
Specimen Preparation ◽  
Cell Shrinkage ◽  
Whole Cells ◽  
Critical Point Drying ◽  
Prepared Material ◽  
Inverted Microscope ◽  
Phosphate Buffered Saline

The use of co-polymerized glutaraldehyde-carbohydrazide (GACH) was proposed for specimen preparation in scanning electron microscopy (SEM) as a means of avoiding dehydration in organic solvents, and to provide dimensionally stable biological specimens through a process of air-drying. It has been assumed that shrinkage of specimens prepared by the GACH technique should be less than that of conventionally-prepared material by critical point drying (CPD). In a previous study, Bell has reported significant shrinkage of whole cells for SEM. This report compares cell shrinkage in GACH and CPD preparations.Fibroblasts from newborn rats were grown on collagen-coated glass cover-slips (with alpha numeric grids etched onto the surface of the coverslips) in Eagle's minimum essential medium + 10% fetal calf serum for 7 d. (3). Using an inverted microscope with phase-contrast optics, micrographs were taken of the cultures in their live state and 1 h. after fixation with 2.5% glutaraldehyde in Dulbecco's phosphate buffered saline (Figs. 1 and 3).

scholarly journals Ultra-Narrow Metallic Nano-Trenches Realized by Wet Etching and Critical Point Drying

Nanomaterials ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 783
Author(s):  
Jeeyoon Jeong ◽  
Hyosim Yang ◽  
Seondo Park ◽  
Yun Daniel Park ◽  
Dai-Sik Kim
Keyword(s):  
Critical Point ◽  
Terahertz Spectroscopy ◽  
High Surface ◽  
Wet Etching ◽  
Drying Process ◽  
Metal Structures ◽  
Critical Point Drying ◽  
Metal Insulator Metal ◽  
Gap Width ◽  
Optical Structure

A metallic nano-trench is a unique optical structure capable of ultrasensitive detection of molecules, active modulation as well as potential electrochemical applications. Recently, wet-etching the dielectrics of metal–insulator–metal structures has emerged as a reliable method of creating optically active metallic nano-trenches with a gap width of 10 nm or less, opening a new venue for studying the dynamics of nanoconfined molecules. Yet, the high surface tension of water in the process of drying leaves the nano-trenches vulnerable to collapsing, limiting the achievable width to no less than 5 nm. In this work, we overcome the technical limit and realize metallic nano-trenches with widths as small as 1.5 nm. The critical point drying technique significantly alleviates the stress applied to the gap in the drying process, keeping the ultra-narrow gap from collapsing. Terahertz spectroscopy of the trenches clearly reveals the signature of successful wet etching of the dielectrics without apparent damage to the gap. We expect that our work will enable various optical and electrochemical studies at a few-molecules-thick level.

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