A Method for Indefinite Storage of Paraffin RibbonsAuthor(s): John M. Legler and Robert M. WinokurSource: Transactions of the American Microscopical Society, Vol. 90, No. 3 (Jul., 1971), pp.363-365Published by: Wiley on behalf of American Microscopical SocietyStable URL: http://www.jstor.org/stable/3225196 .

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SHORTER COMMUNICATIONS

A METHOD FOR INDEFINITE STORAGE OF PARAFFIN RIBBONS

JOHN M. LEGLER and ROBERT M. WINOKUR

Department of Biology, University of Utah, Salt Lake City, Utah 84112

LEGLER, J. M. & WINOKUR, R. M. 1971. A method for indefinite storage of paraffin ribbons. Trans. Amer. Micros. Soc., 90: 363-365. Paraffin ribbons can be stored safely and indefinitely in modified cake boxes (8 X 8 X 3/") with transparent windows. The placing of ribbons on strips of paper facilitates subsequent removal of sections from anywhere in a series. The method of storage is especially valuable when it is impossible to section and mount simultaneously.

Paraffin sections are usually mounted on slides soon after they are cut. There are relatively few techniques for long-term storage of ribbons described in the literature. Immediate mounting of sections is practical only when sufficient time is available for both cutting and mounting or when only a few sections are needed.

In the course of research on the cranial morphology of turtles (NSF Grant GB 16249) we developed a safe and precise system of storing paraffin ribbons for indefinite periods. Serial sections of entire heads were mounted in two or three series (i.e., every second or third section per series), each of which was stained by a different technique. This necessitated the ultimate division of ribbons into individual sections. Technical assistance was provided chiefly by part time students of varying skill. Sectioning and mounting were seldom done by the same person, and it was never possible to section and mount in the same operation. The following method of ribbon storage permitted this operation to work smoothly and at a pace commensurate with the amount of assistance available. We submit that the method could be of value to other workers.

Containers for ribbons were modified from standard, cardboard cake boxes. The boxes can be purchased in quantities of 250 or more from most paper whole- salers. They are sold flat, with creases for folding by hand. Unmodified boxes are eight inches square and four inches deep, with transparent windows in the top and in one side (windowless boxes also available). The flat sheets are cut (Fig. 1) to reduce the depth to % inches. They are then folded, the corners stapled, and the lid hinged with masking tape. The top window remains. Total cost per box is 10-15 cents, including labor.

Paraffin ribbons are placed on 7/," x 112" strips of paper; the strips are assembled on 71/ x 71/" sheets of paper and placed in the boxes (Fig. 2). We use coarse filter paper for the strips and sheets; cheap mimeograph paper works as well and, if dark colored, offers greater visibility of sections. Identifying data are written in soft pencil on the outside of the box.

Most books on microtechnique (e.g., Gray, 1964; Galigher & Kozloff, 1964) imply that sections should be mounted immediately, suggest that storage is harm- ful to ribbons, or ignore the subject of storage completely. Jones (1966) and Humason (1967) have recommended the use of various clothing boxes for storage. Feldman & Harrison (1969) describe a shallow storage device made from a file folder and soda straws.

We tried a wide variety of boxes and other containers before deciding on the box herein described. The obtaining of boxes (e.g., graph paper, hosiery) from TRANS. AMER. MICROS. Soc., 90(3): 363-365. 1971.

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364 TRANS. AMER. MICROS. SOC., VOL. 90, NO. 3, JULY 1971

I

I

I

BOTTOM

FIG. 1. A flat cake box, 25 X 16", as purchased. Size when folded is 8 X 8 X 4". Dashed lines indicate cuts necessary to reduce depth to 3%". Creases for folding are not visible.

FIG. 2. Ribbon box in use. Note that all ribbons can be removed simultaneously or they can be handled by individual paper strips.

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SHORTER COMMUNICATIONS 365 SHORTER COMMUNICATIONS 365

stores was inconvenient and the supply was neither constant nor uniform enough for our needs. Most such boxes are too shallow; air currents produced when the box is opened are likely to disturb thin ribbons. Eight-inch square boxes fit and stack easily on standard shelving. The transparent window facilitates identifica- tion of sections without opening the lid. Use of paper strips permits the individual handling of small parts of a series without exposing other sections to the hazards of air currents and dust. Brief data for one or more sections can easily be written on the strips in pencil.

Sections have been stored in the manner described for two to three years at varying temperatures (65-85 F) and relative humidity (10-80%) without detectable change in consistency or staining characteristics (e.g., H & E, Mallory-Heidenhain- Azan, Bodian Protargol-Gold Chloride). Static electricity in the transparent win- dow has never been a problem; as a precaution we routinely pass an ionizing unit (Staticmaster, Mod. 2U500) over the window before using a new box.

LITERATURE CITED

FELDMAN, M. L. & HARRISON, J. M. 1969. A compact storage tray for paraffin ribbons. Stain Technol., 44: 108-109.

GALIGHER, A. E. & KOZLOFF, E. N. 1964. Essentials of practical microtechnique. Lea and Febiger, Philadelphia. 484 pp.

GRAY, P. 1964. Handbook of basic microtechnique, 3rd ed. McGraw-Hill, New York. 302 pp. HUMASON, G. L. 1967. Animal tissue techniques, 2nd ed. W. H. Freeman, San Francisco.

569 pp. JONES, R. M. 1966. Basic microscopic technics. Univ. Chicago Press, Chicago. 334 pp.

FEEDING ON GASTROTRICHS BY THE HELIOZOON ACTINOPHRYS SOL

EUGENE C. BOVEE and DEAN L. CORDELL

Department of Physiology and Cell Biology, University of Kansas, Lawrence, Kansas 66044

BOVEE, E. C. & CORDELL, D. L. 1971. Feeding on gastrotrichs by the heliozoon Actinophrys sol. Trans. Amer. Micros. Soc., 90: 365-369. Actinophrys sol feeds both solitarily and cooperatively on gastrotrichs, especially Chaetonotus vulgaris. The gastrotrich adheres to axopods, is pulled toward the heliozoon by their contractions, is enclosed by bending of adjacent axopods and by a protoplasmic collar which flows over the "belt" of axopods, all within 15 minutes after capture. One gastrotrich nearly escaped the enclosing collar, but was then engulfed, headfirst, within another 20 min- utes. It was dead within one hour after capture. A pair of Actinophrys simultaneously engulfed a gastrotrich in a single fused- and shared-gastriole. Digestion of this gastro- trich was complete within six hours; the residue was voided from the gastriole, and the two Actinophrys separated.

Evidence that protozoa feed on the smaller metazoa is scanty in the literature, with few recent reports (e.g., Brown & Jenkins, 1962; Janovy, 1963; Loosanoff, 1959). The ciliate Dileptus eats planarians (Brown & Jenkins, 1962), becoming gigantic (Janovy, 1963), on rotifers (Sandon, 1932), and feeds competitively on them, or on injured molluscs, nematodes, oligochaetes, trematodes and their cer- cariae, or on Hydra (Brown & Jenkins, 1962). Another ciliate, Condylostoma, eats

TRANS. AMER. MICROS. SOC., 90(3): 365-369. 1971.

stores was inconvenient and the supply was neither constant nor uniform enough for our needs. Most such boxes are too shallow; air currents produced when the box is opened are likely to disturb thin ribbons. Eight-inch square boxes fit and stack easily on standard shelving. The transparent window facilitates identifica- tion of sections without opening the lid. Use of paper strips permits the individual handling of small parts of a series without exposing other sections to the hazards of air currents and dust. Brief data for one or more sections can easily be written on the strips in pencil.

Sections have been stored in the manner described for two to three years at varying temperatures (65-85 F) and relative humidity (10-80%) without detectable change in consistency or staining characteristics (e.g., H & E, Mallory-Heidenhain- Azan, Bodian Protargol-Gold Chloride). Static electricity in the transparent win- dow has never been a problem; as a precaution we routinely pass an ionizing unit (Staticmaster, Mod. 2U500) over the window before using a new box.

LITERATURE CITED

FELDMAN, M. L. & HARRISON, J. M. 1969. A compact storage tray for paraffin ribbons. Stain Technol., 44: 108-109.

GALIGHER, A. E. & KOZLOFF, E. N. 1964. Essentials of practical microtechnique. Lea and Febiger, Philadelphia. 484 pp.

GRAY, P. 1964. Handbook of basic microtechnique, 3rd ed. McGraw-Hill, New York. 302 pp. HUMASON, G. L. 1967. Animal tissue techniques, 2nd ed. W. H. Freeman, San Francisco.

569 pp. JONES, R. M. 1966. Basic microscopic technics. Univ. Chicago Press, Chicago. 334 pp.

FEEDING ON GASTROTRICHS BY THE HELIOZOON ACTINOPHRYS SOL

EUGENE C. BOVEE and DEAN L. CORDELL

Department of Physiology and Cell Biology, University of Kansas, Lawrence, Kansas 66044

BOVEE, E. C. & CORDELL, D. L. 1971. Feeding on gastrotrichs by the heliozoon Actinophrys sol. Trans. Amer. Micros. Soc., 90: 365-369. Actinophrys sol feeds both solitarily and cooperatively on gastrotrichs, especially Chaetonotus vulgaris. The gastrotrich adheres to axopods, is pulled toward the heliozoon by their contractions, is enclosed by bending of adjacent axopods and by a protoplasmic collar which flows over the "belt" of axopods, all within 15 minutes after capture. One gastrotrich nearly escaped the enclosing collar, but was then engulfed, headfirst, within another 20 min- utes. It was dead within one hour after capture. A pair of Actinophrys simultaneously engulfed a gastrotrich in a single fused- and shared-gastriole. Digestion of this gastro- trich was complete within six hours; the residue was voided from the gastriole, and the two Actinophrys separated.

Evidence that protozoa feed on the smaller metazoa is scanty in the literature, with few recent reports (e.g., Brown & Jenkins, 1962; Janovy, 1963; Loosanoff, 1959). The ciliate Dileptus eats planarians (Brown & Jenkins, 1962), becoming gigantic (Janovy, 1963), on rotifers (Sandon, 1932), and feeds competitively on them, or on injured molluscs, nematodes, oligochaetes, trematodes and their cer- cariae, or on Hydra (Brown & Jenkins, 1962). Another ciliate, Condylostoma, eats

TRANS. AMER. MICROS. SOC., 90(3): 365-369. 1971.

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