Transmission Electron Microscopy: A Surgical Pathology Tool for Neuroblastoma

Summary

Pediatric small round blue cell tumors are an intriguing and challenging collection of neoplasms. Therefore, transmission electron microscopy (TEM) and professional knowledge of pediatric tumors can be extremely valuable in surgical pathology. Here, we present a protocol to perform TEM for diagnosing neuroblastoma, one of the most common solid tumors in childhood.

Abstract

Pediatric small round blue cell tumors (PSRBCT) are an intriguing and challenging collection of neoplasms. Light microscopy of small round blue cell tumors identifies small round cells. They harbor a generally hyperchromatic nucleus and relatively scanty basophilic cytoplasm. Pediatric small round blue cell tumors include several entities. Usually, they incorporate Wilms tumor, neuroblastoma, rhabdomyosarcoma, Ewing sarcoma, retinoblastoma, lymphoma, and small cell osteosarcoma, among others. Even using immunohistochemistry, the differential diagnosis of these neoplasms may be controversial at light microscopy. A faint staining or an ambiguous background can deter pathologists from making the proper diagnostic decision. In addition, molecular biology may provide an overwhelming amount of data challenging to distinguish them, and some translocations may be seen in more than one category. Thus, transmission electron microscopy (TEM) can be extremely valuable. Here we emphasize the modern protocol for TEM data of the neuroblastoma. Tumor cells with tangles of cytoplasmic processes containing neurosecretory granules can diagnose neuroblastoma.

Introduction

The work of a pathologist may be pretty challenging both in clinical diagnostics and research fields. The evolution of light microscopy in the 18^th and 19^th century was remarkable. The power of an electron microscope relies primarily on the wavelength of the electrons, which is shorter than light^1,2,3. Before the advent of the polyclonal and monoclonal antibodies and their application in immunohistochemistry, TEM played an influential role in diagnosing small round blue cell tumors.

Starting with the 90's of the last centu....

Protocol

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. The electron microscopy study is part of the normal routine for microscopic investigation of the samples received for diagnostic purposes and does not require approval by the Bioethics Committee. This study is retrospective and respects the complete anonymity of.......

Discussion

In this narrative review with associated protocol, we highlighted the distinctive ultrastructural features of the neuroblastoma. In the end, we suggest that electron microscopy is far from a "dead" or ancient technique, postulating the discovery of a new role if combined with single-cell omics technologies. This contribution would like to emphasize the never ancient role of electron microscopy in pediatric pathology^4,27. The critical steps, the modificati.......

Materials

Name	Company	Catalog Number	Comments
Acetone, ACS Reagent	Electron Microscopy Sciences	10014	Reagent as established by the American Chemical Society (ACS Reagent).
Automated tissue processor	Electron Microscopy Sciences	L12600	LYNX II Automated Tissue Processor for Histology and Microscopy.
Digital camera software	Gatan	L12600	Digital Micrograph
Spurr's resin	Electron Microscopy Sciences	14300	Embedding resin. It provides excellent penetration for embedding tissues and rapid infiltration. The blocks have excellent trimming and sectioning qualities, while thin sections reveal tough qualities under the electron beam.
Ethyl Alcohol	Electron Microscopy Sciences	15058	100% Ethyl alcohol with molecular sieve, 50% and 70%.
Glutaraldehyde, 25% EM Grade Aqueous in Serum Vial	Electron Microscopy Sciences	16214	2.5% gluteraldehyde in 0.1 M cacodylate buffer, pH 7.2-7.4 made from 25% gluteraldehyde, primary fixative for TEM tissue specimens.
Osmium tetroxide	Electron Microscopy Sciences	19110	Second fixative during TEM tissue processing used as OsO4 in distilled water
Polyethylene capsules	Electron Microscopy Sciences	70021	Flat Bottom Embedding Capsules, Size 00
Scintillator	Electron Microscopy Sciences	82010	Phillip Quad Detector
Single Edge Razor Blade	Electron Microscopy Sciences	71952-10	Blade for Clean Rm., 10/Disp. .
Sodium Cacodylate Buffer	Electron Microscopy Sciences	11655	Sodium Cacodylate Buffer, 0.4M, pH 7.2, prepared from Sodium Cacodylate Trihydrate
Tannic Acid, Reagent, A.C.S., EM Grade	Electron Microscopy Sciences	21700	Reagent as established by the American Chemical Society (ACS Reagent).
Transmission Electron Microscope (1)	Hitachi	Hitachi 7100	We use it at the HV 75 setting
Transmission Electron Microscope (2)	JEOL	JEM-1010	We use it at the HV 38 setting
Toluene	Electron Microscopy Sciences	22030	Reagent as established by the American Chemical Society (ACS Reagent)
Ultracut microtome	Leica	11865766	Ultramicrotome
Uranyl acetate	Electron Microscopy Sciences	22400	Uranyless, substitute for uranyl acetate