In the age where early diagnosis of disease is of vital importance we need quick and reliable diagnostic methods with minimal turn around time.

The need for minimum turnaround time has encouraged newer techniques of staining in field of cytology , which require lesser time and are cost- effective without compromising on the cell morphology.[1]

This has led to invention of stain that require lesser time without compromising reliability in diagnosis, leading to innovation of modified ultra fast pap stain.

Quick diagnosis in cytology is the key to a successful treatment and it plays an important role in prognosis of the disease.

The papanicolaou stain is a nucleo-cytoplasmic staining technique developed by George N Papanicolaou, the father of cytopathology in 1942 and subsequently modified by him in 1954 and 1960.

Papanicolaou (Pap) staining method has revolutionized cytology and remains the gold standard in cytology staining. Papanicolaou stain is the preferred stain for cytology since the beginning.

Several modifications have been developed in the pap stain to improve the staining quality &/or to minimize staining time.[2]

To overcome limitations of routine Pap staining several modifications were proposed and an ultrafast papanicolaou stain was introduced by yang and Alvarez in 1994-95 which is a hybrid of romanowsky stain and pap stain.

It not only reduces the time for pap stain to 90 seconds, but also enhances the quality.

It involves rehydration of air dried smears, fixation in alcoholic formalin and subsequent pap staining except that the duration of each step is shortened.[3]

This technique was further modified as many of the reagents were not easily available in our country AS RESULT- Modified Ultra Fast Papanicolaou stain (MUFP) WAS INVENTED BY dr.kamal MM, Bodele A, Munshi MM, Bobhate SK, Kher AV FROM Department of Pathology, Govt. Medical College, Nagpur-2000.

Objective

The objective of our study was to assess the feasibility and applicability of Modified Ultra-Fast Papanicolaou stain (MUFP) with that of routine Papanicolaou stain in – fine needle aspiration of various organs in comparison to standard Papanicolaou stain.

FNA for early diagnosis.

cytology of fluids

A total of 50 specimens were collected from different organs and their comparison was made. thyroid(10), Breast(10), lymph nodes(10) Ascitic fluid (6) Soft tissue swellings(lipoma)-(4) and salivary glands(10) Two sets of smears were made and stained with routine Pap and MUFP with patient’s clinical details. All smears were compared in 6 parameters and Quality index was calculated. EVALUATION OF STAINING QUALITY The stained smears were examined and compared for quality of staining method employed in terms of fixation, nuclear staining, cytoplasmic staining, drying artifacts, overall staining and background of the smear. Each criterion was graded and scored individually as 1, 2 and 3. 1 being minimal score or poor grading and 3 being maximum score or excellent. Benign thyroid follicular cells obscured by blood (Pap); Follicular cells in clean background (Modified Ultra-Fast Pap (MUFP) FIBROADENOMA OF BREAST   TUBERCULOUS LYMPHADENITIS PLEOMORPHIC ADENOMAOF PAROTID ASCITIC FLUID Lipoma

Table No. 1: Correlation of Background in PAP, & MUFP

Table No. 2: Correlation of Overall staining in PAP, & MUFP

Table No. 3: Correlation of Cell Morphology in PAP, & MUFP

Table No. 4: Correlation of AIR DRYING ARTIFACTS in PAP,& MUFP

MUFP stained smears showed clean background and less air drying artifacts. The alcoholic formalin used in MUFP takes lesser staining time for fixation and makes nucleoli to appear red and prominent.

MUFP stain was found to be best for all inflammatory lesions and also for lymph node lesions.

MUFP stain - Lymph Node > Thyroid / Salivary Gland > Breast

PAP stain - Thyroid/ Breast > Lymph Node > Salivary Gland

(> MEANS BETTER THAN)

The advantages of modified Ultra-Fast pap stain (MUFP) compared to Papanicolaou stain 

Staining solution can be prepared from locally available reagents.

Replacing Gill’s hematoxylin with Harris hematoxylin does not alter the staining characteristics and gives equally good results.

As fixation is not required, the staining time is 90-120 seconds and therefore very useful for intraoperative cytology, rapid assessment of adequacy of samples and rapid diagnosis AND MASS SCREENING.

Background is clear, RBC free and thus helps in better interpretation. This is especially useful for smears of vascular organs like the thyroid AND HEAMORRHAGIC SMEARS.

The technique provides good nuclear and cytoplasmic details as the cells appear large with crisp morphological features.[4]

Air drying removes the artifactual changes seen in wet fixed smears due to poor fixation.

The technique causes no deleterious effect on immunophenotyping.

Cell loss with wet fixation is avoided, and therefore recommended for lipid rich tumors like lipoma.

The disadvantages of modified Ultra-Fast Pap stain (MUFP) compared to Papanicolaou stain

• complete air drying should be strictly observed. Inadequate drying gives suboptimal results. Further, smears need to be properly prepared as thick smears don’t give satisfactory results. [5]
• Interpretation of cytoplasmic keratinization is not possible due to the omission of Orange-G.
• Normal saline, Harris hematoxylin and EA-36 should be changed regularly.
• Bipolar single nuclei are not stained properly.
• Locally available solutions may influence the results adversely.

The solution is storage sensitive and the pH OF alcoholic formalin should be maintained at 5.0.

Although pap stains better than mufp; Lesser staining time along with unequivocal morphological quality is undoubtedly the need of the hour for any cytopathology set-up. MUFP stain easily fulfills these criteria either equivalent to or better than the CONVENTIONAL Pap technique for cytological staining and study of various organs. MUFP stain is fast, reliable and can be done with locally available reagents, and therefore is especially useful in developing countries WHICH HAVE LIMITED RESOURCES like India. It causes no effect on immunophenotyping. Cell loss with wet fixation is less, and recommended for lipid rich tumors. The stain was useful in clearing the haemorrhagic background, mucin was better stained and nuclear morphology was highlighted. Hence, MUFP can be included as route stains in cytopathology.

1. 2nd International Conference on Cytopathology & Histopathology- August 10-12, 2016 Las Vegas, USA.
2. Modified ultrafast Papanicolaou staining technique: A comparative study- Moni Thakur, Venkateswara Rao Guttikonda-JOURNAL OF CYTOLOGY-2017
3. Comparison of modified ultrafast Papanicolaou stain with the standard Papanicolaou stain in cytology of various organs-Dr. Anjali Patrikar, Dr. Archana Joshi, Dr. Vidula Govardhan, Dr. Trupti Dongre - International Journal of Scientific and Research Publications, March 2016
4. Comparison of modified ultrafast Papanicolaou stain with the standard rapid Papanicolaou stain in cytology of various organs- Priyanka Choudhary, S Sudhamani, Ajita Pandit, and VM Kiri –RESERCHGATE- 2012

BIOLAB DIAGNOSTICS LAB MANUAL OF PAP AND MUFP STAINS.