Modern hair restoration surgery involves moving hair-bearing grafts from the posterior scalp into areas of hair loss for correction of androgenic alopecia. Over the past decade, this procedure has been refined in multiple ways for the creation of increasingly natural results. The following is a comparison of commonly used graft harvesting, sectioning, and implantation techniques currently in use for hair restoration today. This comparison includes data on total procedure time, graft cutting time, graft insertion time, labor requirements, and cost.

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An existing technique (Vindelov et al. Cytometry 3:323, 1983) has been modified for DNA flow cytometry of human epidermis obtained from 2 to 3 mm punch biopsies. By varying the length of time of digestion of the epidermal disc by trypsin from 5 to 70 Key terms: Trypsin digestion, biopsy, keratimin a controlled release of keratinocytes occurred beginning with the stratum basale and proceeding toward, but not including, the superficial layer of the epidermis, the stratum corneum. nocyte, isolated nucleiAn important prerequisite for performing flow cytometry (FCM) is the acquisition of a single cell or nuclear suspension. For epidermis, the production of single cell suspensions permits the study of a variety of cell types such as Langerhans cells, keratinocytes, keratinocyte subpopulations, and basal cells (2-11). Because of our interest in the possibility of using cell cycle kinetic data obtained from human epidermis as indicators of the best circadian time for maximal host tolerance to chemotherapy, i.e., chronochemotherapy (11, we modified a n existing technique (12) for use in DNA FCM of human epidermis.Dispersion of epidermal cells or their nuclei into suspension has been accomplished by several different methods including trypsin, dithiotheritol (DTT), EDTA, ultrasonication, or ultrasonication plus DTT (2). The trypsin detergent method described by Vindelov et al. (12) has the advantages of (a) frozen storage of the specimen, (b) production of clean nuclei after digestion of cytoplasmic proteins with trypsin, and (c) quantitative staining of DNA with propidium iodide.This report describes a modification of an existing technique (12) for the production of isolated nuclei of keratinocytes obtained from 2-3 mm punch biopsies of adult human epidermis. Based on the duration of the incubation time in trypsin, a controlled release of keratinocyte populations can be achieved. Trypsin digestion dissolved epidermal cell cohesion beginning in the basal cell layer with progression toward the superficial-most layer after longer incubation. MATERIALS AND METHODS BiopsiesThe skin biopsies used in this study were obtained from the scalp during hair transplantation surgery. Epidermal-Dermal Separation and Freezing of Isolated EpidermisTo facilitate the removal of epidermis from dermis, skin biopsies were first individually placed in 0.5% cold acetic acid for 48 h. With the aid of a dissecting microscope, the epidermis was separated from the dermis using watchmaker's forceps. The disc of epidermis was then placed in fresh DMSO solution and frozen rapidly to -70" C. Intact biopsies as well as the peeled dermis and epidermis were stained with hematoxylin and eosin (H & E) and examined by routine light microscopy. Trypsinization and StainingThe trypsin-detergent method of Vindelov (12) was used with the modification that digestion times in trypsin were varied from 5 to 70 min. In addition to varying the digestion time, other variables which were studied included mechanical vortexing at 1 min, 3 min and 5 min intervals during th...

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