Monday, 23 February 2015

Salimetrics Europe publishes new Catalogue, e mailed free of charge upon request

The Salimetrics Europe Team have just put together a new catalogue in pdf format











This 30 page Catalogue is a great teaching aid for University departments running courses in Psychology, Psychiatry, Endocrinology, Reproductive Biology, Sports Science, Diagnostics and Animal Welfare


A copy will be sent to you free of charge by e mailing europe@salimetrics.com

Here is a Preview:







Tuesday, 10 February 2015

Updated Version: Salimetrics One World: Profiles of International Leaders in Salivary Bioscience: Dr Nicolette (Lettie) Bishop, Reader in Exercise Immunology, Loughborough University

Each month we will feature an expert from the Salimetrics Saliva Research Community. The intention is to bring together University Researchers from around the World in order to encourage the sharing of ideas












Updated Biography: Dr Lettie Bishop, Loughborough University


Nicolette (Lettie) Bishop completed her first degree in Sport and Exercise Science at the University of Birmingham in 1997 for which she was awarded the C.T.M Davies Prize for Outstanding Academic Achievement.
Following this, Lettie accepted a PhD Scholarship to work with Professor Mike Gleeson investigating nutritional influences on the immune response to exercise – she continues to work with Mike at Loughborough University, where she first became a Lecturer in Exercise Physiology in September 2000, becoming Senior Lecturer in 2007 and was appointed to her current position in the School as a Reader in Exercise Immunology in December 2014.
Lettie has been PI and Co-I for grant funding of over £2 million and is a collaborator in the Leicester-Loughborough Diet, Lifestyle and Physical Activity Biomedical Research Unit, co-supervising a study into the benefits of regular exercise in patients with chronic kidney disease (CKD).  She has published over 80 peer-reviewed journal articles, editorials, book chapters and review articles in the area of exercise immunology, including as a co-author on the Feb 2011 International Society of Exercise and Immunology Position Statement on Immune function and Exercise.  Since July 2013 Lettie has been an Associate Editor for Medicine and Science in Sports and Exercise, with responsibility for manuscript submissions in the area of exercise, immune function and inflammation.

Lettie has developed her research programme from a keen interest in the effects of exercise on immune cell function and infection risk in elite athletes to a sustainable programme of internationally recognised research investigating the impacts of exercise on systemic and mucosal immunity, chronic systemic inflammation and infection in those with specific long-term conditions.  Her recent research is investigating the relationships between an active lifestyle and markers of immune function and chronic inflammation in patients with Chronic Kidney Disease with colleagues of the Leicester Kidney Exercise team at University Hospitals Leicester. Their work in this area was highly commended at the Da Vinci Health Technology awards in March 2010 and the Loughborough University Enterprise Awards in 2014 and is part of the Leicester/Loughborough Diet, Lifestyle and Physical Activity Biomedical Research Unit (BRU).  Lettie also has a strong interest in translating knowledge from healthy individuals to populations with impaired immune function and as such is currently investigating novel systemic and mucosal immune markers to assess the impact of physical activity on immune and inflammatory status in healthy adults and those with other chronic conditions associated with impaired immune function and chronic inflammation.  She still retains her interests with elite sport, with recent and on-going collaborations with the English Institute of Sport and the England and Wales Cricket Board.

Interview with Lettie

1. Can you tell us about the major themes in your research program?
Effect of exercise on the systemic and mucosal immune system and how this relates to infection risk in the athlete population and also in the general population. Role of exercise on immune function and infection risk in active and inactive individuals with chronic conditions including spinal cord injury and chronic kidney disease.

 2. If you had to pick 1 publication in the past 5 years as the "best of your best", what would it be and why? In terms of saliva research, probably Leicht CA, Bishop NC & Goosey-Tolfrey, VT  (2011) Mucosal immune responses to treadmill exercise in elite wheelchair athletes. Medicine and Science in Sports Exercise 43:1414-1421.  This was the first paper of our work in wheelchair athletes with tetraplegia, paraplegia and those without spinal cord injury (SCI) but with other disabilities such as amputation.  By far the majority of research in the area of exercise immunology has focused on able-bodied athletes and with no real attention paid to mucosal immune responses in wheelchair athletes. Some of these individuals have high level spinal cord injuries that can affect their sympathetic nervous system (SNS), and as a result, alter their mucosal immune response to exercise. However, despite missing central control, these wheelchair athletes show responses thought to be governed by the SNS, such as reductions of saliva flow rate as a result of strenuous exercise. Similarly, we have recently published data showing that chronic exercise responses in athletes with high level SCI are comparable to able-bodied athletes (Leicht et al., International Journal of Sports Physiology and Performance, epub Dec 2011) ,  as decreases in markers of immune function during periods of heavy training have been reported in both populations. It therefore appears that the loss of central control of effector organs via the SNS may be compensated in some way, perhaps via enhanced spinal reflex activity, adapted parasympathetic nervous system activity, or increased sensitivity of receptors involved in autonomic pathways.

3. How did you get interested in using saliva in your research?
During my PhD I began using saliva measurements of immune function because they reflect the level of protection against airborne pathogens, like cold viruses, more than investigations of individual circulating immune cell functions.  Saliva is also easy to collect in any situation, be in in the lab, on the training field or a training camps.


4. Which salivary analtyes are you working with? We work with IgA, cortisol, alpha amylase, lactoferrin, lysozyme, testosterone and chromagranin A

5. How has working with saliva changed the direction of your research plans?Recently my work on saliva immune responses has given me the opportunity to work with colleagues at the Peter Harrison Centre for Disability Sport here at Loughborough  and through this, wheelchair athletes and non-athletes as it gives a reliable marker of mucosal immune protection in a population of people for whom respiratory infections are common .  Collecting saliva has the advantage of being convenient to collect in both laboratory studies and field studies.  This work has now led to collaborations with research groups working with individuals with SCI in both the Netherlands and the United States.

6. What analyte is not measured in saliva now that you would hope could be measured in the future?A reliable biomarker for autonomic nervous system activation - α-amylase is often used as a biomarker of SNS activation, even though correlation of α-amylase and sympathetic markers, such as circulating adrenaline and noradrenaline, are relatively small.  We have found that amylase responses to exercise are similar between those with and those without high level SCI, despite the disruption to the SNS in those with tetraplegia so it isn't a suitable marker in this population.  

7. What advice would give young investigators who might be considering working with saliva in their research? Make sure you are aware of all of the factors that can influence saliva secretion (chewing/sight/smell of food, tooth brushing, time of day, method of collection, how you report your values, e.g relative to albumin, saliva volume, etc ) before you start - there are so many factors that can affect the secretion rate and composition of the secreted saliva it is important that you make sure you control for this as much as possible,

8. Tell us something about you (a hobby or special interest) that we would be surprised to know? For the past 9 years Lettie has somehow juggled working 'part-time' with looking after to her two young boys - how she manages this remains a mystery to her but she wouldn't have it any other way and she is definitely proud of her new found skills in creative birthday cake making - all thanks to them! 



To Contact Lettie e-mail: N.C.Bishop@lboro.ac.uk

Tuesday, 20 January 2015

Researching Sleep? take a look at the Salimetrics 4hour Salivary Melatonin Assay, the most efficient, most precise and easiest to use protocol commercially available


The Salimetrics™ Salivary Melatonin Assay Kit is a competitive immunoassay specifically designed and validated for the quantitative determination of melatonin in saliva for research or circadian/biorhythm assessment. 








The Salimetrics propriety ELISA format allows melatonin assay results in four hours or less with a single sample and one wash step. Salimetrics salivary assay kits are expertly designed, developed and validated to ensure accuracy in saliva and proven to deliver precision results for salivary biomarkers. Our specifically designed antibody promises precise and accurate results in less time. 

Melatonin (N-acetyl-5-methoxytryptamine) is a compound secreted mainly by the pineal gland, but synthesized also in many other tissues and cells. In humans, nocturnally peaking oscillations of melatonin are involved in sleep-wakefulness where melatonin concentrations are lower during the day. In recent years, the role of melatonin and its metabolites have been identified as potent, broad acting antioxidants and free radical scavengers in addition to a playing a role in the upregulation of antioxidant enzymes. 

Melatonin levels in plasma are paralleled by corresponding variations in saliva where the saliva concentrations are about 30% of that found in plasma. Measurement of salivary melatonin is advantageous, especially to avoid invasive venipuncture procedures

View Assay and Assay Protocol (Including competitor assay comparisons)

Wednesday, 14 January 2015

Planning to set up a Saliva Testing Laboratory at your Institution, here is the ultimate wish list for "Gold Standard Results" (Using Salimetrics Industry Leading Assays of course!)



So your research involves the collection and testing of Saliva Samples

Perhaps you are looking to measure Cortisol, Alpha Amylase e.t.c.

Link to our Full Product Range and Testing Protocols

Here we help you understand the required equipment you will need in your laboratory, however don't fret, we have a fully equipped Robotic Platform at our Cambridge UK lab to process samples from within the UK/Europe, we also have extensive laboratory capability within the USA and Australia with a number of independent Centres of Excellence around the Globe

Below we describe the equipment installed in our Cambridge, UK Lab where we also offer Spit Camp intensive training courses


Basic Laboratory
You will need to have a general laboratory lab space within which to conduct your Research. For example.....
  • Laboratory benching, that meets the current specifications for work with Saliva
  • Adequate ventilation
  • Precision balances (For weighing saliva samples to determine flow rate and weekly checks on your pipettes)
  • Vortex mixers (useful for mixing reagents or making saliva dilutions)
  • Heater/ stirrers (useful for mixing wash buffers)
  • Single and multichannel pipettes (essential - these are used for accurately dispensing saliva and reagents into your assay kit. The quality of the final assay results is greatly influenced by pipetting - both in terms of operator experience and well maintained pipettes)

Air conditioning (to keep lab temperature to around 21C) Most Salimetrics assays are conducted at room temperature but if the lab is much colder or warmer than 21C the assay results may be unreliable
The assays most commonly use aspirate/ dispenses of:
Single pipette: <10µl & 25µl & 50µl
Multichannel:  100µl, 200µl and 320µl  (for salivary Alpha Amylase)

High Quality Water
As a minimum you need grade 3 distilled water but ideally ultra-pure water (18 MΩ water) would be used for all buffers in ELISA and salivary Alpha Amylase. Essential to use: Grade 3 distilled a minimum specification. We currently use a Purite Neptune for ultra pure water (18.3 MΩ water) and a Purite Select for the grade 3 water.

Centrifuges:      
The rate-limiting step for our assays is often how quickly we can get samples centrifuged.  Most samples require 1500RCF for 15 minutes and do not require temperature control. As the saliva is being centrifuged it is important to use sealed adaptor lids so in the event that a tube breaks in the centrifuge, saliva is not passed into laboratory air.

Class 11 Cabinets
The class II cabinets are used to protect both the operator and contents of the cabinet from bacterial/ viral infection.  Whether or not a class II is required depends on the institutional view on health & safety and the nature of the samples being analysed. Not essential but the decision to install depends on the Health and Safety policy of your institution. Given that the Salimetrics laboratory processes large and continuous numbers of samples from a wide number of European customers and participants, we chose to use class11 cabinets for all our testing. We use cabinets by Bioquell and Faster
We use twin HEPA filtration but given the work we do there is little need for ways to sterilize the cabinet or filters (for example UV Lights) As you may be operating equipment such as washers, inside cabinets it is useful to have electrical sockets installed in the cabinet

Refrigeration
Many reagents need to be stored at 2-8ºC (fridge) and saliva samples, ideally, at -80C or below.  Fridges should be high quality laboratory specifications that can maintain a stable temperature.  (‘Domestic’ fridges often show large temperature fluctuations between 2 -8ºC), this is essential
We largely use Gramm Fridges and -20C freezers and New Brunswick -86ºC freezers
Ambient fridges can maintain 22ºC, which is very useful if your lab temperature is not always well controlled. not essential but we use an LMK ambient incubator

Wireless Temperature Monitoring
It is very useful to get 24/7 warnings of when fridges/ freezers fail, for example during power cuts. Not essential but if you have to consider the costs of replacing kits or unique saliva samples it may be well worth the cost of monitoring. There are a number of solutions depending on the size of your lab and the how and when you wish to be contacted. We are currently using a system from Tutela medical (http://www.tutelamedical.com)

Liquid CO2 Injection for -80c Freezers
If the temperature of the -80ºC freezer falls below a certain predefined temperature liquid C02 is injected into the freezer to keep the contents cold
Not essential but provides an additional amount of time before you start to risk damage to unique samples. Currently we use a single C02 bottle can maintain our primary -80ºC freezer at -70C for around 6 hours and additional bottles can extend this.  This system provides some ‘peace of mind’ during power cuts or system failures.

We currently use a New Brunswick C02 backup system.
If you are using C02 liquid you will also need a C02 sensor and alarm as the CO2 Gas can be fatal  at <5% concentration in air. (I.e. in case of a C02 gas leak).  We use Quantam sensor and alarm system

Microplate Reader (Essential)
Simple absorbance reader required with most common filters being 405nm (for salivary Alpha Amylase)), 450nm (for most ELISA) and 620nm (for reference read) we currently use a Tecan Sunrise and a Tecan Infinite F200.  The Sunrise is an entry-level system that is more than adequate for Salimetrics ELISA.
It is important to get some form of data processing software with the reader, for example Tecan Magellan software.
If you are considering salivary Alpha Amylase assays then an incubator option is desirable.

Plate Washer (Essential)
8 or 16 channel head will be sufficient, we currently use the Tecan Hydroflex with 18 channel head and 3 reagent bottles. If you are doing more than a few ELISA plates the washer is all but essential.

Plate Shaker Incubator
An orbital shaker that can ideally shake at least two plates at the same time.  If you were considering non Salimetrics ELISA or salivary Alpha Amylase then an incubator unit would be very useful. Not essential but relatively inexpensive and whilst you can tap plates to mix  a shaker provides far more standardised and efficient shaking

LIMS (Laboratory Information Management System)
LIMS databases store, analyse and report assay data.  They are useful if you are processing large numbers of samples, The database also helps to ensure you have detailed information on the location and history of each sample that is required for storage under the Human Tissue Act. We use a custom LIMS from AIS LIMS ltd (www.ais-lims.com)

Automated Liquid Handler
Automated Liquid Handlers can partially or totally automate the ELISA process, but far from essential. All the Salimetrics assays were developed to be used in laboratories without automated equipment. However if you are looking to test large numbers of samples there are advantages in using an automated system. We use a Tecan Freedom Evo with 8 fixed tips, fast wash system, robotic manipulator, 2 x incubator units (MIOs), Tecan Sunrise and Tecan Hydroflex controlled using TecanEvoWare and Tecan EvoWare PLUS software with Liebert UPS power backup.  The unit is enclosed in a Bigneat HEPA filter cabinet. These installations has just been replaced so we now have some 10 years experience of this assay and have just invested in the latest automated liquid handler from Tecan Robot available (There are other manufacturers but this was our first choice)

Autoclave (Essential)
The autoclave disinfects all lab waste and saliva by sterilizing at 131ºC for 30 minutes.  Normal lab waste, once sterilized, goes into the normal waste system and saliva is then incinerated. We use a benchtop 40L LTE Osprey autoclave

Human Tissue Act License
The handling and storage of Saliva falls within the Human Tissue Act. We therefore have invested in an HTA License costing some £6,000 per annum and have been audited by representatives from the HTA and received full approval for our documentation and systems

Salimetrics Saliva Testing Service

As you can see the Salimetrics testing laboratory in Cambridge has state of the art equipment and staffed by colleagues with over a decade of experience in saliva testing. We pride ourselves  by offering a fast, efficient and accurate service to researchers across Europe. Testing, dependent upon the number of samples,  can be as low as £8 per duplicate sample (all inclusive, Assay, Kits, Collection Devices, Bar Code Labelling, Shipping Cool Box) and in addition to the published range of Assays we will always look at other Analytes, so just ask

We also offer guidance on Saliva Collection advice, always talk to us first to avoid mistakes! We can help with project planning and continued customer service support throughout your project

Need more advice, got a question e mail europe@salimetricseurope.com 

Or call +44 (0) 1638 782619