An Inexpensive Vibrating Microtome for Sectioning Fixed Tissue

A tissue sectioner which uses a vibrating razor blade and a simple mechanism for the elevation of the tissue can be constructed for less than fifty dollars. The razor blade is fixed to the vibrator of a hair clipper and a nut and bolt serve as the tissue advance mechanism. A metal disc attached to the nut is used for the stage. The tissue advance mechanism is placed inside a machined teflon cylinder which provides a smooth surface upon which the razor assembly is moved. Fixed tissue may be sectioned uniformly at a thickness of 50 μm or more. This device has the advantages of portability, rapidity of sectioning, and inexpensive construction.     

A Rapid Celloidin Method for the Rotary Microtome

A method is described which combines the writer's hot celloidin technic¹ with a form of the clearing-before-cutting procedure. The method requires only 16-17 days and yields a block which may be cut in any microtome, the sections being as thin as those afforded by paraffin with comparable material. The advantages of celloidin over paraffin, listed in the writer's earlier paper, are retained in the present method which, altho consuming more time than the hot process, requires less skill and gives superior results.     

A Thermoelectrically Cooled Microtome Table and Knife

Thermoelectric cooling units (Frigistor thermoelements) have been used to replace CO₂ gas and solid CO₂ for microtome stage and knife cooling. These units consist of assemblies of series-connected thermoelements, functioning by the Peltier effect. Cooling is controlled by the direct current supply to the units. Current supply is from a double power pack giving 15 amp at 4.8 v for the knife cooling units, and 15 amp at 1 v for the stage. By varying the current flow, the optimum cutting temperature can be obtained and held indefinitely. An 8-couple Frigistor unit replaces the CO₂ stage of a Lipshaw freezing microtome. The stage temperature may be lowered to -36 C in 40-60 sec and at the optimum cutting temperature, 5 μ serial sections of fixed frozen tissue are obtainable. Four 12-couple units are used to cool a 160 mm Jung plane wedge microtome knife fitted to a Reichert sledge microtome, with the stage cooled by one 8-couple unit. The knife temperature can be lowered to -20 C in 5 min; the stage in 1 min. The apparatus has been used to cut a variety of unfixed rat and mouse tissues. The optimum sectioning temperature for such unfixed liver, kidney, spleen, lymph glands, heart, testes, small intestine, pancreas, skin and lung was -20 to -22 C.     

Vitreous Carbon: A New Material for Making Microtome Knives

The quality of sections obtained by microtomy depends to a large extent on the quality and characteristics of the microtome knife itself. Despite the need for improved microtomy techniques, there have been few significant developments since the introduction of glass and diamond knives in the 1950's. The manufacture of microtome knives from vitreous carbon provides new possibilities for developing both improved methods and improved equipment for specimen sectioning. Vitreous carbon has unique physical properties that lend themselves to the generation of precision cutting edges. Such an edge can be obtained either by breaking a piece of vitreous carbon or by using lapidary techniques. The resultant edge seems well adapted to both thick and thin sectioning. The introduction of vitreous carbon as a sectioning tool offers a significant alternative to metal, glass and diamond knives.     

Thermoelectric Freezing for Attaching Nitrocellulose Blocks to the Microtome Stage

The routine production of large sections and of a few sections from a large number of blacks of tissue embedded in nitrocellulose often presents difficulties, large blocks often loosen if cemented to an intermediate carrier in the routine manner.     

Thermoelectric Freezing for Attaching Nitrocellulose Blocks to the Microtome Stage

The routine production of large sections and of a few sections from a large number of blacks of tissue embedded in nitrocellulose often presents difficulties, large blocks often loosen if cemented to an intermediate carrier in the routine manner.     

Glass Knife Adapter for Cutting Epoxy Sections on a Conventional Rotary Microtome

Electron microscopy-style fixation followed by epoxy plastic embedding is often now the method of choice for preparing tissue even for light microscopy; I have found it excellent for fluorescence, autoradiographic and conventional histology (Shaw 1972, 1977). Sections more than about five microns thick can be cut on a really sharp steel knife if the plastic is reasonably soft (Stretton and Kravitz 1973, Shaw 1972), but this is much easier and knife marks are reduced if extra-wide glass knives are used on a special-purpose intermediate microtome like the Sorvall JB-4. Recent budgetary restrictions made us defer purchase of such a microtome, and some alternative had to be devised. I report here a simple but rugged adapter for glass knives which replaces the steel knife in a conventional Leitz rotary microtome and allows thin plastic sections to be cut as easily as with a more sophisticated cutter. It could be adapted for any rotary microtome, and can be readily constructed in most machine shops for negligible cost.     

Microtome sectioning causes artifacts in rhizobox experiments

Experimental data on rhizosphere characteristics at high spatial resolution are required to improve our knowledge on phytoavailability of nutrients and pollutants. In numerous studies, sectioning using refrigerated microtomes has been employed to obtain thin soil layers at defined distances from the root surface. In this study, we assessed the effect of thin slicing and freezing on soil chemical characteristics. Two experimental soils were frozen at –20°C and sliced using a refrigerated microtome. In general, chemical changes relative to the non-sliced control were more pronounced as the trim thickness (thickness of a single slice) decreased. Maximum increases in pH and electrical conductivity (EC) for the smallest trim thickness used (20 m) were 0.9 units and 50%, respectively. Extractable fractions of P (0.5 M NaHCO₃) K, Mg, Mn, Na and Si (1 M NH₄NO₃) increased up to 40, 91, 19, 621, 50 and 100%, respectively. Based on these results, we suggest to use a trim thickness of 200 m. Apart from slicing, freezing (a prerequisite for the microtome technique) was found to bias soil chemical parameters. To circumvent microtome-related artifacts we present a home-made slicing device as a cost-effective alternative, which allows sectioning of non-frozen rhizosphere soil employing one single slice.     

Paraffin Compression Due to the Rotary Microtome

The extent of compression of microtome sections has been studied for blocks with tissue and also blocks of clear paraffin. Thick sections are commonly compressed 15% or more, while in sections below 5 or 10 μ, compression may exceed 50%. Compensatory thickening of sections occurs. The degree of compression for various paraffin samples and for various conditions of knife edge, temperature, etc., is compared. Microscopical work, particularly where quantitative data or reconstructions are involved, is often seriously unpaired by unrecognized artifacts of sectioning. The present work indicates the magnitude of such artifacts. Compensation for distortions of sections is not easy because tissues, particularly dense tissues, may compress less than the paraffin matrix. Section corrugation is due to this inequality in compression. Absorption of water in section flattening causes some tissue readjustment, but this varies with different tissues and different fixations.