Use of Peldri II (a fluorocarbon solid at room temperature) as an alternative to critical point drying for biological tissues

Kennedy, John R.; Williams, Richard; Gray, John

doi:10.1002/jemt.1060110205

Cited by 44 publications

(23 citation statements)

References 13 publications

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“…The use of HMDS is also a safer and cost-effective alternative to CPD. Several previous studies have shown that the use of HMDS yields comparable results as those obtained with identical biological specimens or sample tissues prepared with CPD (Bray et al, 1993, Hochberg and Litvaitis, 2000, Kennedy et al, 1989, Nation, 1983, Slizova et al, 2003, Ubero-Pascal et al, 2005. The data presented here demonstrate that application of HMDS to LVSEM sample preparation is appropriate for preserving and drying mammalian eggs and early zygotes.…”

supporting

confidence: 72%

Ultrastructural analysis of egg membrane abnormalities in post-ovulatory aged eggs

Dalo¹,

McCaffery²,

Evans³

2008

Int. J. Dev. Biol.

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Microscopic analyses of mammalian gamete ultrastructure have provided among the most seminal insights into fertilization. Here we present a summary of ultrastructural studies of mammalian fertilization, together with a review of the effects of post-ovulatory aging in eggs, and our own results using scanning electron microscopy to examine the effects of post-ovulatory aging on the egg membrane topography. Our previous work detected two abnormalities in egg membrane function in aged eggs: aged eggs appeared to be less able to support sperm-egg membrane interaction, thus rendering the eggs less fertilizable, and aged eggs appear to have a reduced ability to prevent polyspermy at the level of the egg membrane, i.e., to establish a membrane block to polyspermy. In the work presented here, we tested the suitability of both environmental (ESEM) and low-vacuum (LVSEM) modes of the FEI Quanta 200 Environmental Scanning Electron Microscope to study mammalian gametes. While ESEM mode was not sufficient under the conditions we used to observe fine cell surface details, the LVSEM mode proved to be an excellent way to view egg membrane topography. Unfertilized aged eggs have an abnormally distended amicrovillar region over the meiotic spindle, and fertilized aged eggs had three different abnormalities detected, from reduced sperm-induced membrane remodeling to abnormal sperm-induced membrane remodeling and membrane blebbing. Combining these insights of egg membrane function and topography with others in the field of post-ovulatory aging, we have expanded insights into why fertilization at increased times after ovulation is associated with poor reproductive outcomes.

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supporting

confidence: 72%

Ultrastructural analysis of egg membrane abnormalities in post-ovulatory aged eggs

Dalo¹,

McCaffery²,

Evans³

2008

Int. J. Dev. Biol.

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“…One of these methods (Kennedy et al, 1989) is similar to freeze drying and utilizes a sublimation dehydrant, Peldri 11, which is frozen and then sublimed from specimens. The Peldri 11 procedure requires no special equipment but can take several hours to perform a run.…”

Section: Introductionmentioning

confidence: 99%

Comparison of hexamethyldisilazane (HMDS), Peldri II, and critical‐point drying methods for scanning electron microscopy of biological specimens

Bray

Bagu

Koegler

1993

Microscopy Res & Technique

245

153

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Three different drying methods, critical-point drying (CPD), Peldri II, and hexamethyldisilazane (HMDS), were compared using representative animal (rat kidney, trachea, duodenum, lung, and red blood cells) and plant (leaves from ten species of monocotyledons and dicotyledons) specimens. All three drying methods produced identical results with animal specimens. Plant specimens showed signs of shrinkage regardless of which drying method was employed. The order of preservation quality from best to worst for leaves was CPD > Peldri II > HMDS, with the CPD method providing substantially better results in all but one case. Postfixation of leaves with osmium tetroxide resulted in poorer preservation in all instances. Peldri II caused complete extraction of leaf cuticular wax, while both both CPD and HMDS showed minimal extraction compared with samples air dried directly from acetone. These results indicate that HMDS provides a time-saving and inexpensive alternative to CPD for animal specimens. Plant specimens, particularly those containing cells with large central vacuoles, are adequately preserved only with the CPD method. In addition, postfixation with osmium should be avoided when processing plant specimens for scanning electron microscopy.

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“…Distortion of tissue surfaces as a result of surface tension produced by different evaporation rates is the major artifact of this technique. Peldri I1 (Ted Pella, Inc., Redding, CAI, a fluorocarbon compound, sublimed from the specimen under vacuum instead of air dried has been used with large pieces of tissue as well as cell suspensions (Helinski et al, 1990;Kennedy et al, 1989). Preservation of tissue structure was reported to be the same as with CPD.…”

Section: Introductionmentioning

confidence: 99%

Scanning electron microscopy of piliated Neisseria gonorrhoeae processed with hexamethyldisilazane

Dekker

Lammel

Brooks

1991

J. Elec. Microsc. Tech.

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Piliated Neisseria gonorrhoeae are virulent and attach readily to some human mucosal cells. The study of interactions between piliated Neisseria gonorrhoeae and surface structures of eukaryotic cells in tissue culture requires consistent high resolution imaging in scanning electron microscopy (SEM). The combination of the fixatives glutaraldehyde, osmium, tannic acid, and uranyl acetate improves preservation of pili and other delicate structures. Following the critical point drying (CPD) process, pili bundles remained intact, but charging produced image distortion in most of the specimens. The use of hexamethyldisilazane (HMDS) with air drying substantially reduced charging and image distortion. Less contrast and greater resolution of pili bundles and surface structures of bacteria or tissue culture cells were obtained at magnifications of 10,000 or higher. As an alternative to CPD, HMDS processing of cell culture monolayers was simple and was more efficient when a large number of samples was processed.

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Use of Peldri II (a fluorocarbon solid at room temperature) as an alternative to critical point drying for biological tissues

Cited by 44 publications

References 13 publications

Ultrastructural analysis of egg membrane abnormalities in post-ovulatory aged eggs

Ultrastructural analysis of egg membrane abnormalities in post-ovulatory aged eggs

Comparison of hexamethyldisilazane (HMDS), Peldri II, and critical‐point drying methods for scanning electron microscopy of biological specimens

Scanning electron microscopy of piliated Neisseria gonorrhoeae processed with hexamethyldisilazane

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