Department of Anatomy, Xuzhou Medical College, Xuzhou, Jiangsu, China
Lymphatic vessels are difficult to be found in the embalmed cadavers during anatomical dissection as they have a transparent thin wall; however, if contrast media have been perfused, they become visible. In the early stages, there were only two major injection techniques for lymphatic studies in unembalmed cadavers, direct and indirect injections. In the former, mercury was utilized as an injectant for identifying lymphatic vessels, but it was abandoned due to its toxicity afterwards. In the latter, the mixture of Prussian blue, turpentine and ether was the injection media for the study, but satisfactory results were not always obtained. Other materials such as coloured dye, iodine or milk and so on were used for injection, which were described in many previous articles and will not get into detail here.
Details of materials and methods for obtaining results in this book are described in the following.
The investigation was performed with appropriate institutional ethics approval. Acquired from deceased people who had donated their bodies, unembalmed (fresh) cadavers were separated to parts that were kept in a deep freezer. Each part of the cadaver was then ready for lymphatic study.
1.2 Materials for Injection
Hydrogen peroxide (6%) (Orion Laboratories Pty Ltd., Australia; Zhonglian Chemical Co., Ltd., Suzhou, China) or mixed with Indian ink (Encre de chine, Pebeo, France) or Prussian blue ink (Code 134010.318 Emb22278J, Le Géant des Beaux-Arts, Saverne Cédex, France) was applied for finding the lymphatic vessels in the tissues. The ratio was 20:1. Both inks did not leak from the vessels into the surrounding tissue if they were injected in lymphatic vessels. Lead oxide mixture was utilized as an injectant for revealing lymphatic vessels. It contained lead oxide (Pb3O4) (Ajax Chemicals P/L NSW Australia), milk powder (Heinz Ltd., Qingdao, China) and water.
Methods for mixture: Put 5 g of milk powder into a pestle and mortar with 3–5 drops of water and mix them well; add 3–4 ml of water to dilute it; add 15 g of lead oxide, mix well and then put in 15 ml of water; mix them very well for use (Fig. 5.1).
A lead oxide mixture
As lead oxide contains a low level of toxic material, it could be replaced by barium sulphate (Shanghai Silian Industry Co. Ltd., China) (Fig. 5.2).
A barium sulphate mixture
1.3 Instruments and Equipment
Instruments include general and microsurgical instruments, such as scalpels, scissors, forcipes, vascular clips, different sizes of needles, syringes and connecting tubes.
If the diameter of the lymph vessel was smaller than 0.3 mm, then a glass needle was used for injection. The glass needle is made of a micro glass tube (Micro Pipets: Fisher Scientific, Pittsburgh, PA, USA) pulled by a glass tube puller (Model PP-830: Narishige Co., Tokyo, Japan) (Fig. 5.3).
Micro glass tubes and glass tube puller
A micromanipulator (MN-153, Narishige Scientific Instrument Laboratory Tokyo, Japan) was used for holding and adjusting the needle during the injection (Fig. 5.4).
A micromanipulator for holding the needle
Surgical microscope (Zeiss, Nano Technology Systems Division, Oberkochen, Germany), microscope (Olympus BH2, Tokyo, Japan), radiological diagnostic machines and accessories (Model Diamond-150TH High-Frequency X-ray Generator, North American Imaging Inc., Camarillo, CA, USA; Fuji FCR IP Cassette: Fujifilm Corporation, Tokyo, Japan; Fuji Computed Radiography Processor: Fujifilm Corporation, Tokyo, Japan), computer (Optiplex 745, Dell, Australia), software (FCR Cr Console Lite, Fuji Photo Film Co., Ltd., Tokyo, Japan), digital camera and so on were utilized for obtaining images, data processing and analysis.
Two steps were included in the technique for finding and perfusing lymphatic vessels. Then specimens were radiographed and photographed for final results. In some cases, a CT scan and histological study were included for acquiring additional information.