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Dr. Pradip Chaudhari
Officer In Charge: Dr. Pradip Chaudhari

This facility focusses on preclinical animal imaging and research on radiopharmaceuticals. The diagnostic radionuclides - technetium-99m and fluorine-18 complexes are being evaluated for their utility in imaging and monitoring cancer xenografts in mouse models. In 2017, the facility supported 17 preclinical PET, SPECT and CT imaging studies on rodents for basic and translational research projects from ACTREC, other DAE units, academic institutes and pharmaceutical industries.

A majority of the studies were proof-of-concept studies, normal tracer uptake studies and in vivo tumor uptake studies - some involving validation of liver and brain xenograft and orthotopic models. The facility also designs imaging protocols for ex vivo bone imaging and analysis utilizing high resolution microCT, development of animal models, data quantitation, and analysis. During 2017, the facility conducted a major imaging study for the pharmaceutical industry. The facility also runs a complete cancer care program for the diagnosis and treatment of pet animals suffering from spontaneous cancer through its animal oncology clinic. During 2017, 122 referral cases underwent major/ minor surgeries, single or combination drug chemotherapy, radiation therapy or a combination of these, based on clinical requirement.

The animal cancer biorepository maintains biological material (blood, fresh-frozen/ formalin-fixed/ FFPE tissues) obtained during diagnosis and treatment, and utilizes them for research on comparative aspects of animal and human cancers. The senior faculty organized a CME on ‘Preclinical imaging and drug discovery’ at ACTREC from 20th to 22nd September 2017. The facility accepted six trainees during the year – one for Bachelor’s and four for Master’s dissertation, one for research experience, and one observer. Orientation tours of the facility were conducted during the year for visiting students and other visitors.

Dr Rajiv Sarin
Dr Rajiv Sarin

The Next Generation Sequencing (NGS) facility at ACTREC has a HiSeq 1500 from Illumina and an Ion Torrent Personal Genome Machine from Thermofisher. The machines were used by in house scientists as well as those from BARC and IISER, Pune during 2017.

Transcriptome analysis was carried out on the Hiseq 1500 to understand the expression of genes in oral cancer cell lines, in human samples exposed to low dose radiation, and in medullary thyroid cancer.

Exome sequencing was carried out to profile mutations in individuals exposed to radiation, for de novo sequencing of an insect exome, and to find differences in the penetrance and disease aggression of germline mutations in inherited cancers. After DNA/ RNA sample quality checks, exome and transcriptome libraries for 55 samples were prepared in house and sequenced on the HiSeq 1500. Good quality data was obtained and downstream bioinformatics analysis was done with open resource tools by the users.

Ion Torrent PGM libraries for targeted resequencing of BRCA1 and BRCA2 genes were prepared in 108 cases of hereditary breast and ovarian cancer syndrome. Pathogenic mutations were identified in 45 families.

MSF
Facility In Charge: Mr. Shashadhar Dolas

Overview

The Mass Spectrometry facility at ACTREC houses four state-of-the-art mass spectrometry platforms connected to high performance liquid chromatography systems – (1) Orbitrap Exploris 480, Thermo, (2) ESI/MALDI Q-TOF Model Synapt XS (3) Triple Quadrupole Model Xevo TQ Absolute and (4) The Nano-LC (ABSCIEX, Eksigent)-ESI-Q-TOF Model Triple TOF 5600 plus SCIEX.

Mass spectrometry is a technique which measures mass-to-charge ratios of ions. This can be used to identify and quantify the molecules of interest in the given sample. Since the advent of soft ionization methods, large biomolecules like proteins are increasingly being analyzed by mass spectrometry for studying various aspects of their structure. Mass spectrometry based Proteomics, i.e., large scale identification and quantification of proteins from a complex mixture of biological origin; and variations thereof, has become an important workflow in biological research. High resolution mass spectrometry is primarily applied for large scale discovery studies while low resolution mass spectrometry is more suited for targeted proteomics. At ACTREC, we have three high resolution mass spectrometers (MS) and one low resolution MS dedicated for omics applications.

Instruments
Cell
Cell

Orbitrap Exploris 480, Thermo

This high resolution MS system has an ESI ion source and a nanoFlex ion source and the mass analyzer is an Orbitrap. This system is coupled with a Vanquish nanoLC. The high resolution and high sensitivity provided by this system is useful in shotgun proteomics applications of discovery nature. There are specialized data analysis software suits for analysis of the raw data, namely Proteome Discoverer and Spectranaut. This system is well known for in-depth proteomic profiling of the sample, label and label-free quantification of the proteins in a complex mixture. This is well suited for comparative proteomics studies leading to biomarker identification for various disease conditions etc.

Cell
Cell

Synapt XS, Waters

This is a high resolution MS with an interesting option of changing the ion sources from ESI, nanoESI and MALDI (matrix assisted laser desorption ionization). For ESI and nanoESI application, it can be coupled with the M-Class UPLC from Waters and can be used for various shotgun proteomics, lipidomics, metabolomics applications. The main mass analyzer is TOF and the system is equipped with Ion Mobility Separation of the ions, providing separation between molecules of same molecular weight but different conformations. Since this system also has an option of Electron Transfer Dissociation (ETD), this is suitable for identification and characterization of the post-translational modifications (PTMs). With MALDI as an ion source, this system can be utilized for tissue MALDI imaging. The data analysis software available is Progenesis QIP and MassLynx.

Cell
Cell

Xevo TQ Absolute, Waters

This is a low resolution MS system with three quadrupole mass analyzers in series. It is coupled with an H-Class UPLC and has an ESI ion source. This system is best suited for targeted proteomics studies, where absolute or relative quantification of the protein/peptide/small molecule of interest can be done using single or multiple reactions monitoring technique (SRM/ MRM) and its variations. The unparalleled sensitivity provided by the MRM assay can be used to track extremely low amounts of compounds of interest in the given sample. The analysis software MassLynx software helps in analyzing the data of targeted quantification experiments

Cell
Cell

TripleTOF 5600 plus, SCIEX

This is a high resolution MS with nanoESI (electrospray ionization) source and nanoLC coupled with it. The main mass analyzer is Time-of-Flight (TOF). This system is useful for shotgun proteomic experiments of qualitative and quantitative nature. The patented SWATH technology of Sciex helps in increasing the depth of the proteomic identification as well as label-free quantification, and thereby able to identify the proteins with very low expression. Along with that, the system can also be used for label-based quantification of proteins. The analysis software ProteinPilot is useful for qualitative and quantitative proteomics studies.

Applications
Sr. No.Applications
1Enriched Proteins (IP / In-gel digested samples)
2Global Protein Profiling (Complex Mixture)
3Label Free Quantification
4Labelled Quantification (i-TRAQ / TMT)
5Parallel Reaction Monitoring (PRM)
6PTMs
7Metabolomics
8Lipidomics
9MALDI MS
10MALDI MS-MS
11Targeted Proteomics Studies
12SRM / MRM studies
Publications

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Staff
Sr. No.NameDesignationEmail ID/Phone/Extension
1Mr. Shashadhar DolasScientific Officer ‘E’ OIC-Mass Spectrometry FacilitySdolas[at]actrec[dot]gov[dot]in
5475/5320
2Dr. Heramb KulkarniScientific Officer ‘D’Heramb.kulkarni[at]actrec[dot]gov[dot]in
5475/5320/5394
3Mrs. Poonam KawleScientific Assistant ‘F’mass.spec[at]actrec[dot]gov[dot]in
5475/5320
4Mr. Prince Ragul V.Technician ’A’mass.spec[at]actrec[dot]gov[dot]in
5475/5320
Contact Details

Mass Spectrometry

KS-08 & KS-19, Khanolkar Shodhika,
Advanced Centre for Treatment, Research & Education in Cancer (ACTREC),
Tata Memorial Centre,
Sector-22, Kharghar,
Navi Mumbai – 410210,
Maharashtra, India.

Email : mass.spec[at]actrec[dot]gov[dot]in
Phone No: +91 (022) 27405000 / 68735000
Extension 5475/5320

Media Gallery
MS
MS
MS
MS

LAF Group
Officer In Charge: Dr. Arvind Ingle

The Laboratory Animal Facility (LAF) is located in an exclusive and specially designed building with over 15,000 sq. ft. area spread over two floors. The first floor houses the rooms for rodents, which are maintained under dark & light cycles and controlled conditions for temperature, humidity, air pressure as per the regulatory requirements with the help of HVAC system. The ground floor houses the service areas (washing, autoclaving and animal feed) and also various laboratories (biochemistry, microbiology, genetic monitoring and histology) and quarantine room.

The Laboratory Animal Facility maintains 10 different basic strains of mice, 1 hybrid strain (B6D2F1) of mice, 10 transgenic mice, 11 knockout mice, and 29 crosses of Tg and knockout mice, one each Nude & SCID mice; one each rat and hamster. Listing of the strains of these animals can be viewed by clicking here- "Strains". Majority of the rodent strains are bred under strictly inbred conditions. The main activity of the Laboratory Animal Facility is to breed, maintain and supply quality rodents to in-house researchers, as per the sanction from the Institutional Animal Ethics Committee (IAEC). The Laboratory Animal Facility is registered with the 'Committee for the Purpose of Control and Supervision of Experiments on Animals' (CPCSEA) for breeding, trade and conducting experiment on small laboratory animals vide registration no. 65/GO/ReiBiBt/S/1999/CPCSEA dt. March 11, 1999. The registration is renewed by the CCSEA till March 2027.

All activities of the Animal Facility are carried out as per standard operating procedures (SOPs). The Animal Facility maintains the records of day to day activities as well as breeding, maintenance and experimentation records of the animals as per the statutory requirement of CPCSEA and keeping in mind the future requirement of the NABL accreditation. Only authorized persons are allowed entry in the animal rooms. Prior approved visitors who have not entered any other outside Animal Facility rooms for last 48 hours, are permitted, if it warrants.

QUALITY CONTROL (QC)

A stringent program for clinico-pathology, microbiology, genetic monitoring of the laboratory rodents and animal feed ensures quality control of the animals. Reporting of the QC tests is done in standard formats and QC reports are maintained in the Animal Facility.

  1. Clinico-Pathology

    For clinico-pathology surveillance, skin/ hair are checked for presence of ecto-parasites. Faecal samples are randomly checked for the presence of endo-parasites by sedimentation method. In addition to sedimentation method, specifically for checking of pinworms infestation, we undertake 'tape test' method on randomly selected mice in all modules of the mice. The weekly sampling schedule is devised such a way that every 12 weeks, representative samples from all the modules are covered under the clinico-pathology. However, the sedimentation and tape tests schedule are maintained a week apart.

  2. Routine/ Conventional Microbiology

    Microbiological examination of animal room air, animal feed, water, bedding material, stool samples and animal organs from moribund animals is also carried out whenever required. The weekly sampling schedule is devised such a way that every 12 weeks representative animal from all the strains are covered under the conventional microbiology testing.

  3. ELISA based Microbiology

    Once in every six months i.e. in February and August every year, randomly selected serum samples are screened for rodent pathogens such as E. cuniculi (EC), Hantan Virus (HV), Lactate Dehydrogenase Elevating virus (LDHV), Lymphocytic Choriomeningitis Virus (LCMV), Minute Virus of Mice (MVM), Mycoplasma pulmonis (MP), Pneumonia Virus of Mice (PVM) and Mouse Hepatitis Viurs (MHV) using ELISA-based ready to use kits. In order to include few new rodent pathogens, the list of the rodent pathogens tested by ELISA methods changes every year.

  4. PCR based Microbiology

    Once in every six months, i.e. in March and September every year, DNA/ RNA samples from randomly selected appropriate tissue samples are screened for fourteen rodent pathogens namely CAR bacilli, Helicobacter bilis, Helicobacter hepaticus, Kilham rat virus, MHV, Murine norovirus, Mycoplasma pulmonis, Pasturella pneumotropica, Pneumocystis carinii, Pseudomonas aeruginosa, Salmonella typhimurium, Sendai virus, Lactate Dehydrgenase Elevating Virus (LDHV), Sendai and Tyzzers organism using polymerase chain reaction (PCR)techniques.

  5. Conventional Genetic Monitoring

    Under the genetic monitoring program, skin grafting and study of biochemical markers for Akp1, Es-1, Es-3, Gpd-1, Gpi-1, Hbb, Idh-1, and Mod- 1 markers are routinely undertaken using cellulose acetate electrophoresis method. Skin grafting tests are conducted in April and October every year. Whereas biochemical markers from randomly selected tissue samples are checked once every six months i.e. in February and August every year.

  6. PCR based Genetic Monitoring

    We also undertake PCR based technique using one each Simple Sequence Length Polymorphism (SSLP) markers from every chromosome as D1Mit21, D2Mit15, D3Mit54, D4Mit53, D5Mit52, D6Mit274, D7Mit77, D8Mit4, D9Mit129, D10Mit10, D11Mit14, D12Mit158, D13Mit3, D14Mit7, D15Mit13, D16Mit4, D17Mit34, D18Mit40 and D19Mit41 for confirmation of the genetic purity of the mice strains. SSLP markers from randomly selected mouse DNA samples are checked once every six months i.e. in May and November every year.

  7. T- and B-cell Profile of Nude, SCID and NSG Mice

    The purity of Nude/SCID/NSG mice breeder as well as experimental animals is routinely checked on the basis of their T- and B- cell profiles using a CD3, CD4, CD8 and CD19 markers by flow cytometry method. Besides, in order to ascertain the immuno-compromised nature of these animals, Animal Facility also randomly checks and track the tumor acceptance in experimental animals of these strains.

  8. Animal Nutrition

    All the species of animals are provided with balanced commercially procured animal feed pellets as per the nutritional requirement of each species/ strain. Microbiological quality of the food is routinely checked in-house whereas nutritional quality of the animal feed is checked by sending the samples to outside testing laboratories.

QUALITY CONTROL SERVICES
ACTREC can provide quality control services of biochemical markers and microsatellite marker tests of genetic monitoring by cellulose electrophoresis and PCR method, respectively as well as ELISA and PCR based tests for the available rodent pathogens to other Animal Facilities in the country on payment basis.
EMBRYO CRYO-PRESERVATION PROGRAM
The LAF has also initiated a program for cryo-preservation of rodent embryos of the available strains of animals in ACTREC. As a part of this programme, embryos from few strains have already been cryo-preserved and revived back to check the efficiency of the cryo-preserved embryos to develop into live animals. This data is also published in reputed international journals.
ICLAS RECOGNITION
Since 2004 the Animal Facility at ACTREC is an Institutional Member of the 'International Council for Laboratory Animal Science' (ICLAS) (http://iclas.org/members/member-list ). Based on efforts of ACTREC in education and training in the field of Laboratory Animal Science, ICLAS have recognized ACTREC Animal Facility as one of the centers in “Training and Education”. This link can be viewed on ICLAS website at http://iclas.org/committees/education-and-training-committee/education. Being institutional member, ICLAS has officially permitted ACTREC Animal Facility to use the ICLAS logo on all certificates issued to the participants attending workshop, certificate courses etc. organized in ACTREC.
NIAW RECOGNISED CERTIFICATE COURSES
ACTREC have conducted two certificate courses on "Laboratory Animal Science" under the MoU with National Institute of Animal Welfare (NIAW), Ministry of Environment, Forests and Climate Change, Government of India. The first batch of 10 participants completed the six-month duration certificate course in Feb. 2013. Second batch of 9 participants completed the 4-week duration certificate course in Dec. 2013. The participants of both the courses received the certificates of participation duly signed by ACTREC and NIAW with official logo of the ICLAS on their certificates.
ORGANISATION OF CONFERENCES/ WORKSHOPS

The Laboratory Animal Facility regularly organizes conferences/ workshops with national and international faculty and participants to keep abreast of the latest developments, evolve regulations and disseminate the knowledge and skills to the laboratory animal professionals within the country. Since 2005, the Laboratory Animal Facility has organized six workshops on various aspects of quality control in ACTREC. We also organized an international conference of LASA India in 2015 in ACTREC

SUMMER / SHORT TERM TRAINING
Laboratory Animal Facility can also induct summer/ short term trainees under the ongoing quality control program of the Animal Facility for the M.Sc. or M.V.Sc. (Part I/II) students whose college requires project work. Interested students may contact Dr. Arvind Ingle, Officer-in-Charge, Laboratory Animal Facility.
ANIMALS FOR RESEARCH
The LAF can provide surplus animals of selected strains to other CPCSEA registered Animal Facilities in India. Request for animals must come from the Officer-in-Charge of the concerned Animal Facility and must contain details of number of animals required, species, strain, age, sex, date on which required, full contact details including e-mail, a copy of the valid/ renewed CPCSEA registration, IAEC approval from the host organization for use of the requested animals, and undertaking that the basic infrastructure for maintaining the required animals is available.

Request for animals, quality control services as well as for summer/ short-term trainings may be addressed to:

Dr. Arvind Ingle, MVSc (Path.); Ph.D., DICVP, FNAVS, FIAVP
Officer-in-Charge, Laboratory Animal Facility
Advanced Centre for Treatment, Research and Education in Cancer
Tata Memorial Centre
Sector 22, Kharghar, Navi Mumbai - 410 210, MS. Tel: 022 - 27405047
Email: aingle[at]actrec[dot]gov[dot]in

Arvind Ingle group
Dr. Arvind Ingle

The Histology facility provides the following services to the Centre: (a) slides of unstained/ haematoxylin and eosin (H&E) stained histology sections of animal tissues including bone/ tumour samples, (b) logistic support for frozen sectioning of human/ animal tissues, and (c) blocks of multiple tissues by pecking method using a microarray machine. During 2017, the facility received 3471 tissue samples in fixative and 1231 human tissue paraffin blocks and, after processing, supplied 6869 stained and 17681 unstained slides to 20 research labs. In addition, 3397 tissues were processed for cryo-sectioning, and 1318 H&E stained and 1318 unstained slides were supplied to 12 research labs.

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