Alternative to xylene as a clearing agent in histopathology

Alwahaibi, Nasar; Aljaradi, Shaima; Alazri, Horiyah

doi:10.4103/jlp.jlp_111_17

Cited by 18 publications

(17 citation statements)

References 12 publications

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“…Therefore, we need to find a substitute for xylene if we wanted to embed large tissues. Various xylene-substitutes had been commercially developed to avoid the most obvious disadvantages of xylene 16,[18][19][20] , including esters, aromatic, naphthenic, higher aliphatic hydrocarbons, terpene-based extracts and chlorinated hydrocarbons. We tested three xylene substitutions and compared them with xylene.…”

Section: Resultsmentioning

confidence: 99%

Paraffin-embedding for large volume bio-tissue

Zhanmu

Yang

Gong

et al. 2020

Sci Rep

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Acquiring ultrahigh-resolution three-dimensional images of large-volume tissues non-human primate tissues was an enormous challenge. Given the preservation of structure and excellent sectioning property, formalin-fixed paraffin-embedding method had an enormous potential for threedimensional reconstruction of fine structures, based on the very thin histological sections and optical images. However, maintaining the structure uniformly in large-volume tissues was difficult during the complex processes. In this study, we presented a detailed protocol for the whole mouse, rat, rabbit brains, and even for the macaque hemisphere. The entire protocol took about 2-30 days to complete for a large sample, including fixation, dehydration, clearing, wax immersion and embedding. In addition, it could be applied to other species and organs, while the embedding processes depended on the size and the type of organs. This method had wide applicability to serve as a baseline for further technique development.

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Section: Resultsmentioning

confidence: 99%

Paraffin-embedding for large volume bio-tissue

Zhanmu

Yang

Gong

et al. 2020

Sci Rep

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“…It is important to note that the blocks in Group E were previously processed with UltraClear™ as mentioned in our previous study. [6] Nuclear and cytoplasm staining, cell morphology, clarity, and uniformity of staining were not well preserved. This might indicate that UltraClear™ processing has affected the efficiency of UltraClear™ in deparaffinization and clearing prior to coverslipping.…”

Section: Discussionmentioning

confidence: 99%

“…All tissues were fixed in 10% neutral-buffered formalin for 24 h. All tissues were processed as previously described using an automated histoprocessor (Spin Tissue Processor Microm STP 120; Thermo Scientific, Walldorf, Germany). [6] One hundred blocks were prepared, and four slides from each xylene-processed blocks and one slide from each UltraClear™-processed block were cut at 3 μm using a rotatory microtome (Leica RM2135, Nussloch, Germany).…”

Section: Methodsmentioning

confidence: 99%

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A substitute to xylene in deparaffinization and clearing prior to coverslipping in histopathology

Alwahaibi

Aldughaishi

2019

J Lab Physicians

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INTRODUCTION: Deparaffinization and clearing prior to coverslipping are important steps in all staining methods in histopathology. Xylene is the most commonly used agent worldwide. However, xylene is toxic. We evaluated safer alternative dewaxing and clearing agents prior to coverslipping in a histopathology laboratory. MATERIALS AND METHODS: Thirteen different fresh surgical tissues were cut into two halves. One half processed using xylene and the other half processed using UltraClear™. Five groups were designed. For each Group of A, B, C, and D, 100 slides were cut from xylene-processed blocks. For Group E, 100 slides were cut from UltraClear™-processed blocks. Group A is the standard method. Group B evaluates UltraClear™ as a dewaxing agent only. Group C evaluates UltraClear™ as a clearing agent prior to coverslipping only. Group D evaluates UltraClear™ as both dewaxing and clearing agents prior to coverslipping. Group E evaluates UltraClear™ as both dewaxing and clearing agents prior to coverslipping. Six parameters were evaluated: nuclear staining, cytoplasmic staining, cell morphology, clarity of staining, uniformity of staining, and cost. RESULTS: Groups B, C, and D showed 79% (P = 0.054), 83% (P = 0.221), and 80% (P = 0.079) adequacy when compared with Group A (89%), respectively. However, Group E showed only 76% (P = 0.016) adequacy. UltraClear™ is more expensive than xylene. CONCLUSION: UltraClear™ is a promising dewaxing agent. It is also a good clearing agent for use prior to coverslipping in histopathology laboratory. Cost-benefit balance between safety of laboratory workers, good quality staining, and cost-effective strategy needs to be further studied.

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“…Staining of wood sections was also done with 1% ethanolic safranin for 10 minutes, distained, counter-stained with fast green, and dehydrated as earlier described. Following dehydration, sections of barks and woods were each cleared in pure xylene for 20 minutes 26 and mounting was done in few drops of Canada balsam.…”

Section: Treatment Of Sections For Microscopic Observationmentioning

confidence: 99%

Evaluation of some pharmacognostic parameters of the herbs for a traditional hematinic (Haematol -B) powdered formulation in Ogbomoso, Nigeria

Ogunkunle¹,

Ideh

Olaniran³

et al. 2021

JPBS

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The traditional hematinic oral powdered herbal drug (Haematol-B) from Ogbomoso, Nigeria had been characterized in terms of its botanical constituents, ascorbic acid and mineral elements composition, but there is inadequate information on the pharmacognostic properties of the ten herbal materials for its formulation. Therefore the objective of this study was to evaluatethe bark and wood anatomy (as appropriate) of seven of these herbs with a view to highlighting the diagnostic markers for their authentication. Following the conventional anatomical procedures of transverse sectioning (TS) and tissue maceration (TIM), a total of 21 characters were drawn from the barks (root or stem as appropriate) of the seven woody species studied, while 41 characteristics of the wood in the roots of three of the species were compiled using TS, transverse longitudinal sectioning (TLS), radial longitudinal sectioning (RLS) and TIM techniques. Staining, mounting and microscopic examinations followed, and the observed features were taxonomically described in accordance with the provisions of International Association of Wood Anatomists. Their diagnostic values among the medicinal herbs studied were also explored. The bark anatomical features that can be used to diagnose the species studied were those of the secondary cortex, phloem rays, axial parenchyma, sclereids and resin ducts. The wood anatomical markers included features of the vessels in the TS and variable ray characteristics in the TLS. The two artificial keys constructed from discontinuities in qualitative and quantitative anatomical features of the barks and the wood are useful tools for avoiding misidentification of the herbal materials studied.

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Alternative to xylene as a clearing agent in histopathology

Cited by 18 publications

References 12 publications

Paraffin-embedding for large volume bio-tissue

Paraffin-embedding for large volume bio-tissue

A substitute to xylene in deparaffinization and clearing prior to coverslipping in histopathology

Evaluation of some pharmacognostic parameters of the herbs for a traditional hematinic (Haematol -B) powdered formulation in Ogbomoso, Nigeria

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