This poster describes the generation of invasive breast cancer cell lines for in vivo imaging, as presented at AACR 2013.
Authors:
Jae Beom Kim, Kenneth Wong, and Konnie Urban (Life Sciences & Technology, PerkinElmer Inc., Alameda, CA).
To find out more about the PerkinElmer technologies presented in this poster, please use the following links:
In Vivo Imaging & Analysis - http://bit.ly/17hNxIE
Microscopy Imaging Systems and Software - http://bit.ly/ZuqpmV
Por que a triagem para SCID?
A Imunodeficiência combinada grave (SCID) é um grupo de distúrbios caracterizados por um defeito grave na produção e no funcionamento da célula. Normalmente, bebês com SCID falecerão por infecções antes de um ano de idade, a menos que o sistema imunológico do bebê seja restaurado através de tratamento [1].
Solução com marca CE para triagem SCID
A PerkinElmer tem orgulho em apresentar o primeiro ensaio comercial para triagem de TREC. O ensaio faz parte de um sistema completo, com marca CE para garantir o uso seguro e eficaz como um auxiliar na triagem de SCID.
A potência do Triplo Quad se combina com a versatilidade do Single Quad.
Limites de detecção incomparáveis, ao nível de ppq. Facilidade de uso inigualável. Finalmente, os analistas podem ter o melhor dos dois mundos com o revolucionário ICP-MS NexION 2000.
O cromatógrafo a gás/espectrômetro de massas (GC/MS) portátil Torion® T-9 da PerkinElmer traz novo significado à portabilidade. Pesando apenas 14,5 quilos, este GC/MS também é rápido, confiável e de fácil utilização.
A PerkinElmer, mantendo seu compromisso com a qualidade e se adaptando sempre às necessidades atuais e futuras, apresenta uma plataforma automatizada que traz imensos benefícios na transferência de líquidos.
Por que a triagem para SCID?
A Imunodeficiência combinada grave (SCID) é um grupo de distúrbios caracterizados por um defeito grave na produção e no funcionamento da célula. Normalmente, bebês com SCID falecerão por infecções antes de um ano de idade, a menos que o sistema imunológico do bebê seja restaurado através de tratamento [1].
Solução com marca CE para triagem SCID
A PerkinElmer tem orgulho em apresentar o primeiro ensaio comercial para triagem de TREC. O ensaio faz parte de um sistema completo, com marca CE para garantir o uso seguro e eficaz como um auxiliar na triagem de SCID.
A potência do Triplo Quad se combina com a versatilidade do Single Quad.
Limites de detecção incomparáveis, ao nível de ppq. Facilidade de uso inigualável. Finalmente, os analistas podem ter o melhor dos dois mundos com o revolucionário ICP-MS NexION 2000.
O cromatógrafo a gás/espectrômetro de massas (GC/MS) portátil Torion® T-9 da PerkinElmer traz novo significado à portabilidade. Pesando apenas 14,5 quilos, este GC/MS também é rápido, confiável e de fácil utilização.
A PerkinElmer, mantendo seu compromisso com a qualidade e se adaptando sempre às necessidades atuais e futuras, apresenta uma plataforma automatizada que traz imensos benefícios na transferência de líquidos.
Como a PerkinElmer contribui para diagnósticos mais rápidos?PerkinElmer, Inc.
Uma doença detectada precocemente, pode acelerar a inicialização de tratamentos eficazes, que mudarão a história de vida de crianças portadoras de doenças raras e, de seu núcleo familiar.
Como a PerkinElmer contribui para diagnósticos mais rápidos?PerkinElmer, Inc.
Uma doença detectada precocemente, pode acelerar a inicialização de tratamentos eficazes, que mudarão a história de vida de crianças portadoras de doenças raras e, de seu núcleo familiar.
Mais do que nunca, tornou uma premissa para os produtores de carnes, Frigoríficos e Laboratórios, garantirem a segurança e a qualidade dos alimentos que produzem.
Saiba como o DA 7250 pode te ajudar a garantir todos os certificados de segurança e qualidade.
O sequenciamento de nova geração (Next Generation
Sequencing - NGS) se tornou uma importante ferramenta
da biologia molecular, dentro das diversas áreas da
pesquisa e do ambiente clínico, devido à capacidade de
sequenciar, ou seja, identificar a ordem dos nucleotídeos que compõe o genoma humano, de forma rápida e em grande quantidade, além de ter reduzido significativamente o custo do sequenciamento por base.
Visando oferecer soluções que atendam aos padrões necessários de qualidade para as reações de NGS e também promover melhoria expressiva no importante processo de construção da biblioteca, a PerkinElmer adicionou ao seu portfólio de Applied Genomics os kits para o preparo de bibliotecas de NGS Bioo Scientific®.
Não importa o tamanho da rotina de seu laboratório. As plataformas LabChip®GX Touch vão se adequar à sua necessidade e proporcionar um fluxo de trabalho muito mais produtivo.
New Research Evaluating Cisplatin Uptake in Ovarian Cancer Cells by Single Ce...PerkinElmer, Inc.
Application Note
Authors:
Lauren Amable
National Institute on Minority Health and
Health Disparities, National Institutes of Health
Bethesda, MD
Stan Smith
Chady Stephan
PerkinElmer, Inc.
Shelton, CT
Cromatógrafo Gasoso PerkinElmer Clarus® All-In-One
Com exclusivo detector FID (Flame Ionization Detector) da família Clarus CG que não requer gás Nitrogênio (Make-Up), sendo necessário para o seu funcionamento apenas os gases Hidrogênio e Ar Sintético, a PerkinElmer mais uma vez inova no setor de Cromatografia Gasosa com sistema CG Clarus All-In-One.
Acontece nesta semana o XXI Encontro de Genética do Nordeste, em Recife (PE). A PerkinElmer e a MultiDiagnóstica apresentam as linhas Chemagen para extração de ácidos nucleicos e LabChip para análises de DNA e RNA.
Analysis of In-Service Oils Following ASTM D5185 with the Avio 200 ICP-OESPerkinElmer, Inc.
With heavy machinery, it is important to assess its status during operation to prevent breakdowns and costly repairs. A key aspect is monitoring the status of the oil or lubricants used to lubricate various components such as engines, transmissions, gearboxes and many other important areas: if the oil degrades too much or becomes highly contaminated, it can damage various components. While many aspects of in-service oils need to be monitored, the metal content provides information about the condition of both the engine and the oil itself to help determine the health of the engine and when the oil needs changing. Because of its importance, ASTM created a method for the analysis of in-service oils: method D5185.
Analysis of Oil Additives Following ASTM D4951 with the Avio 200 ICP-OESPerkinElmer, Inc.
When blending base oils and additives for use as lubricants, it is important to know and control the concentrations of certain
elements for optimal performance and longer engine life. Because this process is so crucial, ASTM developed a method
for monitoring this procedure: ASTM method D4951, which covers barium (Ba), boron (B), calcium (Ca), copper (Cu), magnesium (Mg), molybdenum (Mo), phosphorus (P), sulfur (S), and zinc (Zn). However, not all these elements need to be measured all the time – the blends and additives are specified
to meet certain performance specifications which can vary among oil types, depending on their final use.
A Comparison of Microsecond vs. Millisecond Dwell Times on Particle Number Co...PerkinElmer, Inc.
With the growing popularity of nanomaterials in a wide variety of products and processes, the need to measure and characterize these materials has also grown. For metallic or metal-containing engineered nanoparticles (NPs), single particle ICP-MS (SP-ICP-MS) has been developed and is growing in popularity due to its ability to rapidly detect and characterize a large number of particles, determine particle size and size distributions, the particle number concentration in a sample, and the elemental mass concentration of both dissolved and particulate species. SP-ICP-MS has been applied to a wide variety of sample types1-5, demonstrating its versatility.
A Portaria da Água MS n°. 2914/2011 estabelece os critérios aceitos para que a água seja considerada apropriada para o consumo humano. Dentre os critérios estabelecidos, estão avaliações da presença de compostos orgânicos e inorgânicos, bem como níveis de radioatividade alfa e beta. A PerkinElmer é a única empresa que conta com um portfólio completo para essas análises.
Como a PerkinElmer contribui para diagnósticos mais rápidos?PerkinElmer, Inc.
Uma doença detectada precocemente, pode acelerar a inicialização de tratamentos eficazes, que mudarão a história de vida de crianças portadoras de doenças raras e, de seu núcleo familiar.
Como a PerkinElmer contribui para diagnósticos mais rápidos?PerkinElmer, Inc.
Uma doença detectada precocemente, pode acelerar a inicialização de tratamentos eficazes, que mudarão a história de vida de crianças portadoras de doenças raras e, de seu núcleo familiar.
Mais do que nunca, tornou uma premissa para os produtores de carnes, Frigoríficos e Laboratórios, garantirem a segurança e a qualidade dos alimentos que produzem.
Saiba como o DA 7250 pode te ajudar a garantir todos os certificados de segurança e qualidade.
O sequenciamento de nova geração (Next Generation
Sequencing - NGS) se tornou uma importante ferramenta
da biologia molecular, dentro das diversas áreas da
pesquisa e do ambiente clínico, devido à capacidade de
sequenciar, ou seja, identificar a ordem dos nucleotídeos que compõe o genoma humano, de forma rápida e em grande quantidade, além de ter reduzido significativamente o custo do sequenciamento por base.
Visando oferecer soluções que atendam aos padrões necessários de qualidade para as reações de NGS e também promover melhoria expressiva no importante processo de construção da biblioteca, a PerkinElmer adicionou ao seu portfólio de Applied Genomics os kits para o preparo de bibliotecas de NGS Bioo Scientific®.
Não importa o tamanho da rotina de seu laboratório. As plataformas LabChip®GX Touch vão se adequar à sua necessidade e proporcionar um fluxo de trabalho muito mais produtivo.
New Research Evaluating Cisplatin Uptake in Ovarian Cancer Cells by Single Ce...PerkinElmer, Inc.
Application Note
Authors:
Lauren Amable
National Institute on Minority Health and
Health Disparities, National Institutes of Health
Bethesda, MD
Stan Smith
Chady Stephan
PerkinElmer, Inc.
Shelton, CT
Cromatógrafo Gasoso PerkinElmer Clarus® All-In-One
Com exclusivo detector FID (Flame Ionization Detector) da família Clarus CG que não requer gás Nitrogênio (Make-Up), sendo necessário para o seu funcionamento apenas os gases Hidrogênio e Ar Sintético, a PerkinElmer mais uma vez inova no setor de Cromatografia Gasosa com sistema CG Clarus All-In-One.
Acontece nesta semana o XXI Encontro de Genética do Nordeste, em Recife (PE). A PerkinElmer e a MultiDiagnóstica apresentam as linhas Chemagen para extração de ácidos nucleicos e LabChip para análises de DNA e RNA.
Analysis of In-Service Oils Following ASTM D5185 with the Avio 200 ICP-OESPerkinElmer, Inc.
With heavy machinery, it is important to assess its status during operation to prevent breakdowns and costly repairs. A key aspect is monitoring the status of the oil or lubricants used to lubricate various components such as engines, transmissions, gearboxes and many other important areas: if the oil degrades too much or becomes highly contaminated, it can damage various components. While many aspects of in-service oils need to be monitored, the metal content provides information about the condition of both the engine and the oil itself to help determine the health of the engine and when the oil needs changing. Because of its importance, ASTM created a method for the analysis of in-service oils: method D5185.
Analysis of Oil Additives Following ASTM D4951 with the Avio 200 ICP-OESPerkinElmer, Inc.
When blending base oils and additives for use as lubricants, it is important to know and control the concentrations of certain
elements for optimal performance and longer engine life. Because this process is so crucial, ASTM developed a method
for monitoring this procedure: ASTM method D4951, which covers barium (Ba), boron (B), calcium (Ca), copper (Cu), magnesium (Mg), molybdenum (Mo), phosphorus (P), sulfur (S), and zinc (Zn). However, not all these elements need to be measured all the time – the blends and additives are specified
to meet certain performance specifications which can vary among oil types, depending on their final use.
A Comparison of Microsecond vs. Millisecond Dwell Times on Particle Number Co...PerkinElmer, Inc.
With the growing popularity of nanomaterials in a wide variety of products and processes, the need to measure and characterize these materials has also grown. For metallic or metal-containing engineered nanoparticles (NPs), single particle ICP-MS (SP-ICP-MS) has been developed and is growing in popularity due to its ability to rapidly detect and characterize a large number of particles, determine particle size and size distributions, the particle number concentration in a sample, and the elemental mass concentration of both dissolved and particulate species. SP-ICP-MS has been applied to a wide variety of sample types1-5, demonstrating its versatility.
A Portaria da Água MS n°. 2914/2011 estabelece os critérios aceitos para que a água seja considerada apropriada para o consumo humano. Dentre os critérios estabelecidos, estão avaliações da presença de compostos orgânicos e inorgânicos, bem como níveis de radioatividade alfa e beta. A PerkinElmer é a única empresa que conta com um portfólio completo para essas análises.
micro teaching on communication m.sc nursing.pdfAnurag Sharma
Microteaching is a unique model of practice teaching. It is a viable instrument for the. desired change in the teaching behavior or the behavior potential which, in specified types of real. classroom situations, tends to facilitate the achievement of specified types of objectives.
Flu Vaccine Alert in Bangalore Karnatakaaddon Scans
As flu season approaches, health officials in Bangalore, Karnataka, are urging residents to get their flu vaccinations. The seasonal flu, while common, can lead to severe health complications, particularly for vulnerable populations such as young children, the elderly, and those with underlying health conditions.
Dr. Vidisha Kumari, a leading epidemiologist in Bangalore, emphasizes the importance of getting vaccinated. "The flu vaccine is our best defense against the influenza virus. It not only protects individuals but also helps prevent the spread of the virus in our communities," he says.
This year, the flu season is expected to coincide with a potential increase in other respiratory illnesses. The Karnataka Health Department has launched an awareness campaign highlighting the significance of flu vaccinations. They have set up multiple vaccination centers across Bangalore, making it convenient for residents to receive their shots.
To encourage widespread vaccination, the government is also collaborating with local schools, workplaces, and community centers to facilitate vaccination drives. Special attention is being given to ensuring that the vaccine is accessible to all, including marginalized communities who may have limited access to healthcare.
Residents are reminded that the flu vaccine is safe and effective. Common side effects are mild and may include soreness at the injection site, mild fever, or muscle aches. These side effects are generally short-lived and far less severe than the flu itself.
Healthcare providers are also stressing the importance of continuing COVID-19 precautions. Wearing masks, practicing good hand hygiene, and maintaining social distancing are still crucial, especially in crowded places.
Protect yourself and your loved ones by getting vaccinated. Together, we can help keep Bangalore healthy and safe this flu season. For more information on vaccination centers and schedules, residents can visit the Karnataka Health Department’s official website or follow their social media pages.
Stay informed, stay safe, and get your flu shot today!
Recomendações da OMS sobre cuidados maternos e neonatais para uma experiência pós-natal positiva.
Em consonância com os ODS – Objetivos do Desenvolvimento Sustentável e a Estratégia Global para a Saúde das Mulheres, Crianças e Adolescentes, e aplicando uma abordagem baseada nos direitos humanos, os esforços de cuidados pós-natais devem expandir-se para além da cobertura e da simples sobrevivência, de modo a incluir cuidados de qualidade.
Estas diretrizes visam melhorar a qualidade dos cuidados pós-natais essenciais e de rotina prestados às mulheres e aos recém-nascidos, com o objetivo final de melhorar a saúde e o bem-estar materno e neonatal.
Uma “experiência pós-natal positiva” é um resultado importante para todas as mulheres que dão à luz e para os seus recém-nascidos, estabelecendo as bases para a melhoria da saúde e do bem-estar a curto e longo prazo. Uma experiência pós-natal positiva é definida como aquela em que as mulheres, pessoas que gestam, os recém-nascidos, os casais, os pais, os cuidadores e as famílias recebem informação consistente, garantia e apoio de profissionais de saúde motivados; e onde um sistema de saúde flexível e com recursos reconheça as necessidades das mulheres e dos bebês e respeite o seu contexto cultural.
Estas diretrizes consolidadas apresentam algumas recomendações novas e já bem fundamentadas sobre cuidados pós-natais de rotina para mulheres e neonatos que recebem cuidados no pós-parto em unidades de saúde ou na comunidade, independentemente dos recursos disponíveis.
É fornecido um conjunto abrangente de recomendações para cuidados durante o período puerperal, com ênfase nos cuidados essenciais que todas as mulheres e recém-nascidos devem receber, e com a devida atenção à qualidade dos cuidados; isto é, a entrega e a experiência do cuidado recebido. Estas diretrizes atualizam e ampliam as recomendações da OMS de 2014 sobre cuidados pós-natais da mãe e do recém-nascido e complementam as atuais diretrizes da OMS sobre a gestão de complicações pós-natais.
O estabelecimento da amamentação e o manejo das principais intercorrências é contemplada.
Recomendamos muito.
Vamos discutir essas recomendações no nosso curso de pós-graduação em Aleitamento no Instituto Ciclos.
Esta publicação só está disponível em inglês até o momento.
Prof. Marcus Renato de Carvalho
www.agostodourado.com
CDSCO and Phamacovigilance {Regulatory body in India}NEHA GUPTA
The Central Drugs Standard Control Organization (CDSCO) is India's national regulatory body for pharmaceuticals and medical devices. Operating under the Directorate General of Health Services, Ministry of Health & Family Welfare, Government of India, the CDSCO is responsible for approving new drugs, conducting clinical trials, setting standards for drugs, controlling the quality of imported drugs, and coordinating the activities of State Drug Control Organizations by providing expert advice.
Pharmacovigilance, on the other hand, is the science and activities related to the detection, assessment, understanding, and prevention of adverse effects or any other drug-related problems. The primary aim of pharmacovigilance is to ensure the safety and efficacy of medicines, thereby protecting public health.
In India, pharmacovigilance activities are monitored by the Pharmacovigilance Programme of India (PvPI), which works closely with CDSCO to collect, analyze, and act upon data regarding adverse drug reactions (ADRs). Together, they play a critical role in ensuring that the benefits of drugs outweigh their risks, maintaining high standards of patient safety, and promoting the rational use of medicines.
Best Ayurvedic medicine for Gas and IndigestionSwastikAyurveda
Here is the updated list of Top Best Ayurvedic medicine for Gas and Indigestion and those are Gas-O-Go Syp for Dyspepsia | Lavizyme Syrup for Acidity | Yumzyme Hepatoprotective Capsules etc
Pharma Pcd Franchise in Jharkhand - Yodley Lifesciences
Generation of invasive breast cancer cell lines for in vivo imaging
1. Generation of invasive breast cancer cell lines for in vivo imaging
Jae Beom Kim, Kenneth Wong, and Konnie Urban. Life Sciences & Technology, PerkinElmer Inc., Alameda, CA
Abstract 1 In vivo monitoring of tumor growth 4 Biomarkers in MDA-MB-231-luc2-LN cells
Whole animal non-invasive imaging contributes significantly to understand tumor behavior. It Day 102 Day 107 Day 115 Day 126 Tumor volume Bioluminescence
A
also plays a critical role in drug discovery and development. Optical imaging is convenient
A B
because it does not require radioactive materials for imaging. Especially in preclinical
MDA-MB-231-luc2
M #3 - Excision Day 126
1,000
applications, optical imaging can be a very useful tool because genetic modification is 800
Tumor Volume
600
(mm^3)
feasible. The most popular optical imaging is using bioluminescence. We introduced various 400
La Very Very Large 200
cancer cell lines that express firefly luciferase. These cells have stable expression of light rg Large 0
0 30 60 90 120 150
emission for prolonged cell culture situation. This enables researchers to implant the cells into e Cultured MDA-MB-231-luc2-LN cells were stained with
Experimental Day
Before Unmixing After Unmixing PathScan signaling nodes multiplex
animals and monitor tumor development and metastasis. In addition, these tumor cells can be immunofluorescence kit (Cell Signaling Technology).
detected using fluorescent agents that target tumor cells. As a consequence, one can co- C D Cells were counter stained with DAPI and fluorescent
register both bioluminescent and fluorescent images. Cells also can be labeled with both B Day 73 Day 93 Day 101 Day 106 Tumor volume Bioluminescence Fluorescence images were taken using Nuance camera. Each signal
was spectrally unmixed using Nuance software. (A)
MDA-MB-231-luc2-tdTomato
bioluminescent and fluorescent markers such as luciferases and fluorescent proteins. Image before unmixing, (B) Spectrally unmixed image,
#M6 - Excision Day 108 1.0E+12 M#6 - Excision Day 108 M #6 - Excision Day 108 (C) Unmixed DAPI image, (D) Unmixed Phospho-
Although there are many different types of cell lines available for different tumor types, 3,000 1.0E+11
10
Erk1/2 image (Alexa Flour 488), (E) Unmixed Phospho
Total Radiant Efficiency
9
2,500 1.0E+10 8
Akt (Alexa Flour 555), (F) Unmixed Phospho-S6
Tumor Volume
studying metastasis can be challenging. That is mainly because most cell lines show delayed 7
(photons/sec)
2,000 1.0E+09
Total Flux
(mm^3)
6
( X 10^8)
1,500 1.0E+08 5 ribosomal protein (Alexa Flour 647).
DAPI Phospho-Erk1/2
4
metastasis when implanted in the animal. To expedite the metastasis, intravenous injection or 1,000
500
1.0E+07
1.0E+06
3
2
1.0E+05 1
intracardiac injection is performed to generate secondary tumors in the animal. However, 0
E F
1.0E+04 0
0 30 60 90 120 0 30 60 90 120 150 0 30 60 90 120 150
these methods do not represent true metastasis from originated organs. One of the most Experimental Day M #6 Experimental Day M #6 Experimental Day
Non-Palpable Tumor (N=3) Non-Palpable Tumor (N=3)
popular breast cancer cell line is MDA-MB-231. When these cells are implanted into Background Background
mammary fat pads of female nude mice, it typically takes more than 90 days to detect
MDA-MB-231-luc2 and MDA-MB-231-luc2-tdTomato cells were implanted into the mammary fat pads of athymic nude mice.
metastasis in the secondary sites. Therefore, to study the tumor behavior or to examine the Tumor growth was monitored using bioluminescent imaging. Tumor volumes were measured using a caliper. Bioluminescent
drug efficacy, one has to wait for a long time to see metastasis with MDA-MB-231 cell line. signals were quantitated using Living Image software. Representative mice were shown. (A) Mouse implanted with MDA-
MB-231-luc2 cells. (B) Mouse implanted with MDA-MB-231-luc2-tdTomato cells. Graphs next to the animals images are Phospho Akt Phospho-S6
Here, we generated tumor cell lines that were derived from MDA-MB-231 originated cells. We quantitation for tumor volumes, bioluminescent signals, and fluorescent signals.
took MDA-MB-231 cells that were labeled with either luciferase (MDA-MB-231-luc2) or
luciferase & tdTomato fluorescent protein (MDA-MB-231-luc2-tdTomato). These cells were
implanted into mammary fat pads of nude mice and secondary tumors were isolated from
lymph nodes. Tissues were dissociated to single cells and clonal cell lines were established 2 Tumor metastases with MDA-MB-231-luc2 and MDA-MB-
(MDA-MB-231-luc2-LN and MDA-MB-231-luc2-tdTomato-LN). The growth patterns of these luc2-tdTomato cells and establishment of new cell lines 5 Instruments
cells were compared to their corresponding parental cells. To find out the metastasis patterns
of these cells, we implanted new cell lines orthotopically into nude mice. Our results showed
that these cell lines showed faster metastasis than parental cell lines. Moreover, we examined Spectral Unmixing
Day 107 Day 115 Day 126 Ex vivo Growth Curve IVIS and Nuance camera
biomarker expression patterns with multiplexing multispectral microscopy. These cells can be
MDA-MB-231-luc2
used to study tumor metastasis and drug discovery using non-invasive in vivo imaging. Acquisition
Materials & Methods
MDA-MB-231-luc2
MDA-MB-231-luc2-tdTomato vs. MDA-MB-luc2- RGB
Cell culture
tdTomato-LN
Spectra of labels
-tdTomato
7.00E+05
6.00E+05
Human breast cancer cell line MDA-MB-231 cells were transfected with luciferase 2 cDNA (MDA-MB- 5.00E+05
4.00E+05
Label A Label
3.00E+05 C
231-luc2, PerkinElmer). MDA-MB-231-luc2-tdTomato cells were generated by transfecting tdTomato 2.00E+05
1.00E+05
cDNA into MDA-MB-231-luc2 cells. Cells were grown in MEM media supplemented with 10% fetal 0.00E+00
0 24
MDA-MB-231-luc2-tdTomato
48 72
MDA-MB-231-luc2-tdTomato-LN
96
bovine serum (Hyclone) without antibiotics. Growth curves were generated by seeding 100,000 cells Label B Label
RGB D
in a T25 flask. At each time point, cells were trypsinized and counted using an automatic cell counter Representatio Once unmixed, labels can
MDA-MB-231-luc2 and MDA-MB-231-luc2-tdTomato cells were implanted into the mammary fat pads of athymic nude mice n of Spectral be measured accurately.
Cube
(Nexcelom, Lawrence, MA). Total numbers of cells were plotted in a logarithmic scale. Secondary and tumor metastases were monitored using bioluminescent imaging. Representative mice are shown. Ex vivo images of
Autofluorescence
tumor cell lines were established using limited dilution. The luminescence and/or fluorescence excised lymph nodes and growth curves are shown. Top Row: MDA-MB-231-luc2 cells. Bottom row: MDA-MB-231-luc2-
(removed from quantitation)
tdTomato cells.
stability were monitored for 4 weeks. IVIS Spectrum and Nuance camera are shown. Whole animal in vivo imaging was done using an IVIS Spectrum. Multispectral
fluorescence microscopic images were taken using a Nuance camera. Spectral unmixing for in vivo and in vitro fluorescence
imaging was done using Living Image and Nuance software respectively.
Tumor implantation
All the procedures for animal care and tumor cell implantation followed the approved animal protocols
and guidelines of the Institutional Animal Care and Use Committee (IACUC). Prior to implantation, all
3 Tumor metastases with MDA-MB-231-luc2-LN cells
tumor cells were tested for the presence of mycoplasma and mouse pathogens. Female athymic
Mouse #2 Mouse #5 Mouse #6 Mouse #8 Mouse #9
nude mice (Charles River) were anesthetized with isoflurane and two million MDA-MB-231-luc2 or
MDA-MB-231-luc2-tdTomato cells were orthotopically implanted into mammary fat pads of athymic Summary
nude mice.
1. Tumor growth was monitored using MDA-MB-231-luc2 and MDA-MB-
MDA-MB-231-luc2-LN
Day 24
In vitro and in vivo fluorescent and bioluminescent imaging luc2-tdTomato cells in vivo using bioluminescence and/or
For in vivo imaging, mice were anesthetized with isoflurane. Mice were subjected to in vivo tdTomato fluorescence imaging in real time.
imaging using the IVIS Spectrum (PerkinElmer). Multispectral images were spectrally unmixed using
Living Image software (PerkinElmer). For MDA-MB-231-luc2 tumor imaging in vivo, mice were
injected with D-luciferin. Bioluminescent images were taken using IVIS Spectrum. Once tumor 2. Secondary tumors were isolated from metastasized lymph nodes
metastasized into lymph nodes, secondary tumor was excised from the animals and bioluminescent and new cell lines were established.
images were taken using IVIS Spectrum.
Day 32
3. Newly established cell lines showed different growth patterns from
Histology, Immunofluorescence and tissue image processing
Tissues were harvested at post-implantation day 7 and embedded in paraffin. Sections were cut at 7 those of parental cell lines.
mm thickness. For MDA-MB-231-luc2-LN cells, PathScan Signaling Nodes Mutiplex IF Kit (Cell
Signaling Technology) was used. Staining procedures followed the manufacture’s recommendation.
MDA-MB-231-luc2-LN cells were implanted into the mammary fat pads of athymic nude mice (n=12). Tumor growth was
4. MDA-MB-231-luc2-LN and MDA-MB-231-luc2-tdTomato-LN cell lines
Once the staining was done, cells were mounted using DAPI containing mounting media. Multi- monitored using bioluminescent imaging. Two animals showed no palpable tumor. Six representative animals are shown at day 24
and 32. For imaging, primary tumors were masked to detect metastases in the lymph nodes. Quantitation of bioluminescence
metastasize faster than parental cells.
spectral images were taken with an epifluorescence microscope (Nikon E400) equipped with Nuance
was performed using Living Image software. MDA-MB-231-luc2-tdTomato-LN cells showed similar pattern of metastases (Data
camera (PerkinElmer). not shown).